Oxazinones and methods for their use and synthesis

ABSTRACT

The invention pertains, at least in part, to new intermediates and synthetic methods for the stereospecific synthesis of oxazinone compounds, which are useful, for example, as antibiotics. The invention also pertains to novel olefinic oxazinone compounds, methods for their synthesis, and methods of using these compounds for the synthesis of oxazinones. The invention also pertains to methods for using the compounds to treat bacterial associated states in subjects.

RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional ApplicationSerial No. 60/310,103, entitled “Intermediates and Methods ForSynthesizing Oxazinones,” filed Aug. 3, 2001, the entire contents ofthis application are hereby incorporated herein by reference. Thisapplication is related to U.S. Pat. No. 6,399,600 B1, issued on Jun. 4,2002, the entire contents of this patent are hereby incorporated hereinby reference.

BACKGROUND OF THE INVENTION

[0002] Many antibiotics act by interfering with the biosynthesis ofbacterial cell walls (Strominger et al. J. Biol. Chem. 234:3263 (1959)).The completion of bacterial cell wall synthesis is mediated by enzymestermed penicillin-binding proteins (PBPs) which cross-link differentpeptidoglycan chains. In particular, PBPs link the penultimate D-Alaresidue of a peptidoglycan terminating in a N-acyl-D-Ala-D-Ala moiety tothe terminal amino group of a lysine residue of another peptidoglycanchain. Glycopeptide transpeptidase is an example of a PBP present inmany bacteria.

[0003] Most known PBPs are serine peptidases, which have a conservedSer-X—X-Lys sequence at the active site. The β-lactam family ofantibiotics, whose members include penicillins and cephalosporins,inhibit PBPs by forming a covalent bond with the serine hydroxyl groupto produce an acyl-enzyme. The enzyme is then unable to carry out thefinal step in the biosynthesis of the bacterial cell wall. As a resultthe wall is weakened, becomes permeable to water, and the bacterial cellswells, bursts, and dies.

[0004] The simplest kinetic description of the reaction between abacterial enzyme (Enz) and a β-lactam antibiotic is given in Scheme 1below:

[0005] In addition to the PBP's, many bacteria also produce a secondtype of penicillin-recognizing enzyme, known as a β-lactamase. PBPs andβ-lactamase exhibit the same kinetics as set forth in Scheme 1 above,but with different rate constants. This difference in rate constants hasimportant consequences. In the case of PBP's, k₂>>k₃ (i.e., theformation of the acyl-enzyme is much faster than its hydrolysis). Theresult is that the enzyme is inhibited, and antibacterial activity maybe observed. In the case of a β-lactamase, k₂≈k₃ (i.e., the formationand hydrolysis of the acyl enzyme proceed at comparable rates). Thesekinetics lead to regeneration of the enzyme, and inactivation of theantibiotic as a result of the net hydrolysis of the P-lactam bond in thedeacylation step. The latter sequence of reactions comprises theprinciple mechanism of bacterial resistance to β-lactam antibiotics.Useful antibacterial activity is generally considered to requirek₂/k₁≧1000 M⁻¹ sec⁻¹ and k₃≦1×10⁻⁴ sec⁻¹.

[0006] Resistance to antibiotics is a problem of much current concern.Alternatives to existing antibiotics are invaluable when bacteriadevelop immunity to these drugs or when patients are allergic(approximately 5% of the population is allergic to penicillin). Becauseof the relatively low cost and relative safety of the β-lactam family ofantibiotics, and because many details of their mechanism of action andthe mechanism of bacterial resistance are understood, one approach tothe problem of resistance is to design new classes of compounds thatwill complex to and react with a penicillin recognizing enzyme, and bestable to the hydrolysis step. In order to be effective, theantibacterial agent should have the ability to react irreversibly withthe active site serine residue of the enzyme.

[0007] The crystal structures of β-lactamases from B. licheniformis, S.aureus and E. coli (RTEM) suggest a chemical basis for resistance toβ-lactam antibiotics. Apart from the conserved Ser-X—X-Lys active sitesequence, these β-lactamases have a conserved Glu166 which participatesin the hydrolysis of the acyl-enzyme. It appears that the acylatedhydroxyl group of the active site serine and the carboxyl group of Glu166, together with a water molecule, are involved in the hydrolysisstep. The water molecule and the carboxyl group act in concert and thisinteraction is the source of bacterial resistance to β-lactamantibiotics. Drug design must therefore include a process for theremoval or inactivation of this water molecule.

[0008] Numerous β-lactam compounds have been developed in the past whichare structural analogues of penicillin and can complex to and react withpenicillin recognizing enzymes. Like penicillin, such antibiotics arepresumed to be conformationally constrained analogues of anN-acyl-D-Ala-D-Ala peptidoglycan moiety, the O═C—N β-lactam bond servingas a bioisostere of the D-Ala-D-Ala peptide bond. Effectiveantibacterial activity also requires a properly positioned carboxylgroup or equivalent and a hydrogen bonding hydroxyl or acylamino group.A computer implemented molecular modeling technique for identifyingcompounds which are likely to bind to the PBP active site and, thus, arelikely to exhibit antibacterial activity has been developed (U.S. Pat.No. 5,552,543).

[0009] Some oxazinones having possible biological activity are known inthe prior art Khomutov et al. synthesized tetrahydro-1,2-oxazin-3-one(Chem. Abs. 13754a, 1962) and 4-benzamidotetrahydro-1,2-oxazin-3-one(Chem. Abs. 58, 13944b, 1963). The latter compound is also known asN-benzoyl-cyclocanaline. According to Khomutov, cyclocanaline is knownto inhibit glutamate-aspartate transaminase and exhibits activityagainst tuberculosis bacilli. The structure of cyclocanaline is shown informula (A) below.

[0010] Frankel et al. reported the synthesis of DL-cyclocanaline(4-amino-tetrahydro-1,2-oxazin-3-one) hydrochloride from canalinedihydrochloride in 1969 (J. Chem. Soc. (C) 174601749, 1969) andrecognized that DL-cyclocanaline is a higher homologue of the antibioticcycloserine.

SUMMARY OF THE INVENTION

[0011] The invention pertains, at least in part, to new intermediatesand synthetic methods for the stereospecific synthesis of oxazinonecompounds, which are useful, for example, as antibiotics. The inventionalso pertains to novel olefinic oxazinone compounds, methods for theirsynthesis, and methods of using these compounds for the synthesis ofoxazinones.

[0012] In one embodiment, the invention pertains to olefinic oxazinonesof the formula (I):

[0013] wherein:

[0014] R₁ is an amino acid side chain mimicking moiety;

[0015] R₄, R₆, R₈, and R₉ are substituting moieties;

[0016] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andacceptable salts and esters thereof.

[0017] The invention also pertains, at least in part, to methods forsynthesizing oxazinones of the formula (II):

[0018] wherein:

[0019] R₁ is an amino acid side chain mimicking moiety;

[0020] R₂ is halogen, OH, SH, NH₂, NHCOR₃, or an electronegative moiety;

[0021] R₃ is an antibacterial substituent;

[0022] R₄, R₈ and R₉ are each independently selected substitutingmoieties;

[0023] R₅ is OH, NH₂, NHCOR₃, or an electronegative moiety; and

[0024] R₆ is a substituting moiety or the oxygen of a carbonyl groupwhen taken together with R₅;

[0025] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andpharmaceutically acceptable salts and esters thereof

[0026] The method includes contacting an olefinic oxazinone of formula(I) with a derivatizing agent, under appropriate conditions such that anoxazinone of formula (II) is synthesized.

[0027] In another embodiment, the invention pertains in part to epoxideoxazinones of formulae (III) and (IV):

[0028] wherein:

[0029] R₁ is an amino acid side chain mimicking moiety;

[0030] R₄, R₆, R₈ and R₉ are each independently selected substitutingmoieties;

[0031] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andacceptable salts and esters thereof.

[0032] In yet another embodiment, the invention pertains to methods forthe synthesis of olefinic oxazinones. The method includes contacting adiolefin of formula (V) with a cyclization catalyst under appropriateconditions, such that an olefinic oxazinone of formula (I) is formed.The diolefin of formula (V) is:

[0033] wherein:

[0034] R₁ is an amino acid side chain mimicking moiety;

[0035] R₄, R₆, R₈ and R₉ are each independently selected substitutingmoieties;

[0036] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andacceptable salts and esters thereof.

[0037] The invention also pertains, at least in part, to another methodfor the synthesis of an olefinic oxazinone of formula (I). The methodincludes treating an olefinic triphenyl phosphine salt with ozone underappropriate conditions, such that an olefinic oxazinone is formed. Theolefinic triphenyl phosphine salt is of formula (VI)

[0038] wherein:

[0039] R₁ is an amino acid side chain mimicking moiety;

[0040] R₄, R₆, R₈ and R₉ are each independently selected substitutingmoieties;

[0041] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andpharmaceutically acceptable salts and esters thereof.

[0042] In another embodiment, the invention pertains to a method fortreating a bacterial associated state in a subject. The method includesadministering to said subject an effective amount of an oxazinonecompound of the invention, e.g., a compound of formula (I), (II), (III),(IV) or otherwise described herein.

[0043] In yet another embodiment, the invention includes pharmaceuticalcompositions comprising an effective amount of a compound of theinvention, e.g., a compound of formula (I), (II), (III), (IV) orotherwise described herein.

DETAILED DESCRIPTION OF THE INVENTION

[0044] The invention pertains, at least in part, to new intermediatesand synthetic methods for the stereospecific synthesis of oxazinonecompounds, which are useful, for example, as antibiotics. The inventionalso pertains to novel olefinic oxazinone compounds, methods for theirsynthesis, and methods of using these compounds for the synthesis ofoxazinones.

[0045] In one embodiment, the invention pertains to an olefinicoxazinone of the formula (I):

[0046] wherein:

[0047] R₁ is an amino acid side chain mimicking moiety;

[0048] R₄, R₆, R₈ and R₉ are each independently selected substitutingmoieties;

[0049] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andacceptable salts and esters thereof.

[0050] In a further embodiment, the olefinic oxazinones of formula (I)include olefinic oxazinones represented by formula (VII):

[0051] wherein

[0052] R₁ is an amino acid side chain mimicking moiety;

[0053] R₄ and R₆ are each independently lower alkyl or hydrogen;

[0054] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andacceptable salts and esters thereof.

[0055] The language “amino acid side chain mimicking moiety” includesmoieties that are amino acid side chains or mimic amino acid side chainsand which allow the oxazinone (e.g., a compound of formula II) toperform its intended function by, e.g., mimicking the structure orfunction of an amino acid side chain. For example, the “amino acid sidechain mimicking moiety” allows the oxazinone to interact with the activesite of a penicillin recognizing enzyme. Examples of amino acid sidechain moieties include the side chains of natural and unnatural, D- andL-amino acids. For example, the amino acid side chain mimicking moietymay be the side chain of a neutral amino acid (e.g., alanine, valine,leucine, isoleucine, phenylalanine, tryptophan, or methionine), a polaramino acid (e.g., glycine, serine, threonine, cysteine, tyrosine,asparagine, or glutamine), or a charged amino acid (e.g., aspartic acid,glutamic acid, lysine, arginine, or histidine). In an embodiment, theamino acid side chain mimicking moiety is the side chain of alanine(e.g., methyl).

[0056] In another embodiment, the amino acid side chain mimicking moietyis substituted or unsubstituted alkyl, e.g., lower alkyl. The side chainmimicking moiety may be substituted with any substituent that allows itto perform its intended function (e.g., when present in the oxazinone,it should allow the oxazinone to interact with penicillin recognizingenzyme, etc.). Examples of alkyl amino acid side chain mimickingmoieties include straight chain, branched and cyclic alkyl groups.Examples of alkyl groups include, but are not limited to, methyl, ethyl,i-propyl, n-propyl, i-butyl, n-butyl, t-butyl; pentyl, cyclopentyl,cyclohexyl, or hexyl. Other examples of amino acid side chain mimickingmoieties include alkenyl, alkynyl, carbonyl, aralkyl or aryl moieties.Examples of aryl moieties include substituted and unsubstituted phenyland substituted and unsubstituted heteroaryl.

[0057] The language “protecting moiety” includes groups which can beused to protect the carboxylic acid functionality during synthesis ofthe oxazinone compound. Any protecting moiety known in the art andcompatible with the other functionality of the oxazinones, olefinicoxazinones, and/or epoxide oxazinones may be used. Examples ofprotecting moieties include esters, groups known to those of skill inthe art, and those described in Greene, Protective Groups in OrganicSynthesis, Wiley, New York (1981), incorporated herein by reference.

[0058] The language “prod rug moiety” includes moieties which can becleaved in vivo to yield an active drug (see, e.g., R. B. Silverman,1992, “The Organic Chemistry of Drug Design and Drug Action”, AcademicPress, Chp. 8). Prodrugs can be used to alter the biodistribution or thepharmacokinetics for a particular compound. Examples of prodrug moietiesinclude substituted and unsubstituted, branched or unbranched loweralkyl moieties, lower alkenyl moieties, di-lower alkyl-amino lower-alkylmoieties (e.g., dimethylaminoethyl), acylamino lower alkyl moieties(e.g., acetoxymethyl), acyloxy lower alkyl moieties (e.g.,pivaloyloxymethyl), aryl moieties (e.g., phenyl), aryl-lower alkyl(e.g., benzyl), substituted (e.g., with methyl, halo, or methoxysubstituents) aryl and aryl-lower alkyl moieties, amides, lower-alkylamides, di-lower alkyl amides, and hydroxy amides. Also included aregroups which may not need to be removed to yield an active drug.

[0059] The term “substituting moiety” includes moieties which can beplaced at any one of the R₄, R₆, R₈ or R₉ positions withoutprohibitively detrimentally affecting the synthesis of the antibiotic.In an advantageous embodiment, each substituting moiety is selected suchthat the oxazinone formed may perform its intended function. Examples ofsubstituting moieties include alkyl, hydrogen and other substituentswhich are not detrimental to the synthesis of olefinic oxazinone.Examples of substituting moieties include alkyl (e.g., lower alkyl) andhydrogen. In one embodiment, R₈ and R₉ are both hydrogen.

[0060] In one embodiment, the olefinic oxazinone of formula (I) has theR configuration at the * carbon. In another further embodiment, theolefinic oxazinone of formula (I) has the S configuration at the *carbon as shown in the formulae below:

[0061] In one embodiment, R₇ is hydrogen. In another embodiment, R₄ ishydrogen or lower alkyl (e.g., methyl, ethyl, propyl, etc.). In anotherembodiment, R₆ is hydrogen or lower alkyl (e.g., methyl, ethyl, propyl,etc.).

[0062] The invention also pertains, at least in part to a method forsynthesizing oxazinones of the formula (II):

[0063] wherein:

[0064] R₁ is an amino acid side chain mimicking moiety;

[0065] R₂ is halogen, OH, SH, NH₂, NHCOR₃, or an electronegative moiety;

[0066] R₃ is an antibacterial substituent;

[0067] R₄, R₈ and R₉ are each independently selected substitutingmoieties;

[0068] R₅ is OH, NH₂, NHCOR₃, or an electronegative moiety; and

[0069] R₆ is a substituting moiety or the oxygen of a carbonyl groupwhen taken together with R₅;

[0070] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andpharmaceutically acceptable salts and esters thereof.

[0071] In a further embodiment, compounds of formula (II) includecompounds wherein R₈ and R₉ are each hydrogen, R₄ is hydrogen or alkyl(e.g., lower alkyl) and R₆ is hydrogen, lower alkyl, or the oxygen of acarbonyl group when taken together with R₅.

[0072] The method includes contacting an olefinic oxazinone of formula(I) with a derivatizing agent, under appropriate conditions such that anoxazinone of formula (II) is synthesized.

[0073] The term “derivatizing agent” includes reagents and catalystswhich can be used to derivatize the olefinic bond of the olefinicoxazinone compound. Methods for the derivatization of olefinic bonds areknown in the art (for examples, see, Smith & March, March 's AdvancedOrganic Chemistry, John Wiley & Sons, New York, 2001, and referencescited therein). The term also includes combinations and sequences ofderivatizing agents which form the desired oxazinone through multistepsyntheses from the olefinic oxazinone. For example, in one embodiment,the olefinic oxazinone is contacted with a derivatizing agent underconditions such that an oxazinone epoxide is formed. Then, the epoxideis reacted with another derivatizing agent, under conditions such thatthe desired oxazinone is formed.

[0074] Examples of derivatizing agents include those which selectivelyaminohydroxylate the olefinic bond of the olefinic oxazinone, e.g.,(DHQD)₂PHAL, (DHQD)₂AQN, (DHQ)₂PHAL, (DHQ)₂AQN (see, for example, Tao etal. Tet. Lett. (1998) 39:2507; O'Brien, et al. Angew. Chem. Int. Ed.(1999) 38(3):326); Brunko et al. Angew. Chem. Int. Ed. Engl. (1997)36(13/14):1483). Also included are agents which dihydroxylate theolefinic bond to form the diol. Examples include dihydroquininep-chlorobenzoate and OsO₄ (Wang et al. J. Am. Chem. Soc. (1992)114:7568). Other methods are also well known in the art (see, forexamples, see, Smith & March, March 's Advanced Organic Chemistry, JohnWiley & Sons, New York, 2001, and references cited therein). Otherderivatizing agents include those which selectively halogenate theolefinic bond (e.g., N-fluoropyridium salt (Umemoto et al. J. Am. Chem.Soc. (1990) 112:8563, as well as others known in the art).

[0075] In one embodiment, an oxazinone epoxide is formed when thederivatizing agent contacts the olefinic oxazinone under appropriateconditions. In a second embodiment, an oxazinone epoxide is formed by atwo-step process, comprising contacting the olefinic oxazinone with ahypohalous acid (e.g., HOF, HOBr, HOCl, or HOI), and then subsequentlywith a base (e.g., potassium carbonate).

[0076] Examples of oxazinone epoxides include compounds of formulae(III) and (IV):

[0077] The epoxide oxazinone can be formed using enantioselectiveepoxidation techniques known in the art. For example, the epoxidationcan be performed using the Sharpless asymmetric epoxidation witht-BuOOH, titanium tetraisopropoxide and optically active diethyltartrate included as the derivatizing agents (see, Sharpless et al. PureAppl. Chem. (1983) 55:1823; Pfenninger Synthesis (1986) 89).Alternatively, the epoxide oxazinone can be formed using variousoxidizing agents (e.g., sodium hypochlorite) and optically activemangenese or cobalt catalysts such as manganese-salen or salen (II)cobalt complexes as the derivatizing agents (see, Jacobson et al. J. Am.Chem. Soc. (1991) 113:7063; Hatayama et al. Synlett (1992)407; LinkerACIEE (1997) 36:2060).

[0078] The epoxide oxazinones can then be converted further withadditional derivatizing agents (e.g., epoxide opening agents) underappropriate conditions to yield the desired oxazinone. Examples ofepoxide opening agents include various nucleophiles, water (see, Kotsukiet al. Tet. Lett. (1993) 34:4031; Furrow et al. J. Org. Chem., (1998)63:6776 for stereo- and enantioselective ring opening), ammonia (see,Hou et al. Tetrahedron Asymmetry, (1998) 9:1747; and Meguro J. Chem.Soc. Perkin. Trans. 1, (1994) 2597 for stereo- and enantioselective ringopening), hydrohalic acids (e.g., HF, HBr, HCl, HI) (see, Olah, Isr. J.Chem. (1978) 17:148; Shimizu et al. Tett. Lett. (1988) 29:4101),hydrogenating agents (e.g., SmI₂ (Molander et al. J. Org. Chem., (1986)51:5259), Red-A1 (Gao et al. J. Org. Chem. (1988) 53:4081), H₂ and aPd-phosphine catalyst, etc. (Oshima et al. J. Am. Chem. Soc. (1989)111:6280), etc. These reactions may be performed under conditions suchthat the resulting oxazinone is stereo- and/or enantiospecificallysynthesized. Many methods known in the art for converting epoxides toother chemical functional groups are available (see, for example, Smith& March, Advanced Organic Chemistry, John Wiley & Sons, New York, 2001).

[0079] The term “appropriate conditions” includes the conditions for thevarious chemical reactions which result in the desired product (orintermediate to be formed). For example, appropriate conditions includeschoice of solvent (e.g., polar, non-polar, etc.), atmosphere (e.g., Ar,N₂, etc.), temperature, pressure, etc. One of ordinary skill in the artwill be able to use the techniques described in the application and/orthe chemical literature to determine what the appropriate conditions fora particular reaction are. Furthermore, the desired oxazinone mayrequire additional steps such as purification (e.g., by techniques knownin the art, such as, but not limited to column chromatography,preparative HPLC, distillation, recrystallization, etc.) such that thedesired oxazinone is obtained at the desired purity.

[0080] The term “antibacterial substituent” includes substituents whichare known to enhance the biological activity of penicillins orcephalosporin. Examples of such substituents are given in U.S. Pat. Nos.4,013,653, 4,775,670; and 4,822,788, incorporated herein by reference intheir entirety.

[0081] The term “electronegative substituent” includes substituentswhich are more electron withdrawing than hydrogen. Examples ofelectronegative substituents include carboxylic acids, halogens (e.g.,fluorine, bromine, chlorine, and iodine), aryloxy groups, esters,ethers, ketones, thiols, thiethers, hydroxyl groups, aryl groups,alkenyl groups, etc.

[0082] The invention also pertains, at least in part, to methods for thesynthesis of an olefinic oxazinone of formula (I). One of the methodsincludes contacting a diolefin with a cyclization catalyst underappropriate conditions, such that an olefinic oxazinone is formed. Thediolefin is of formula (V):

[0083] wherein:

[0084] R₁ is an amino acid side chain mimicking moiety;

[0085] R₄, R₆, R₈ and R₉ are each independently selected substitutingmoieties;

[0086] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andacceptable salts and esters thereof. In a further embodiment, R₈ and R₉are both hydrogen.

[0087] The term “cyclization catalyst” includes catalysts known in theart which are capable of cyclizing a diolefin to form a olefinic ring.In one embodiment, the cyclization catalyst is Grubb's Catalyst, e.g.,bis(tricyclohexylphosphine) benzylidene ruthenium dichloride.

[0088] The term “appropriate conditions” include those conditions whichallow the reaction to take place. For example, for this reactionappropriate conditions include those which result in the olefinicoxazinone being formed. Examples of appropriate solvents includenon-polar solvents such as methylene chloride and benzene. However, itshould be noted that the appropriate conditions are reaction specificand an ordinarily skilled artisan will be able to use the teachingsherein as well as the chemical literature, if necessary, to determinethe optimal conditions for his/her particular reaction.

[0089] The invention also pertains to a method for the synthesis of anolefinic oxazinone of formula (I) by treating an olefinic triphenylphosphine salt (e.g., triflic salt) with ozone under appropriateconditions, such that an olefinic oxazinone is formed. The olefinictriphenyl phosphine salt is of formula (VI)

[0090] wherein:

[0091] R₁ is an amino acid side chain mimicking moiety;

[0092] R₄, R₆, R₈, and R₉ are each independently selected substitutingmoieties;

[0093] R₇ is hydrogen, a protecting moiety, or a prodrug moiety, andpharmaceutically acceptable salts and esters thereof. In a furtherembodiment, R₈ and R₉ are both hydrogen.

[0094] The term “alkyl” includes saturated aliphatic groups, includingstraight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, octyl, nonyl, decyl. etc.), branched-chain alkyl groups(e.g., isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl (alicyclic)groups (e.g., cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl), alkyl substituted cycloalkyl groups, and cycloalkylsubstituted alkyl groups. The term alkyl further includes alkyl groups,which can further include oxygen, nitrogen, sulfur or phosphorous atomsreplacing one or more carbons of the hydrocarbon backbone. In certainembodiments, a straight chain or branched chain alkyl has 6 or fewercarbon atoms in its backbone (e.g., C₁-C₆ for straight chain, C₃-C₆ forbranched chain), and more preferably 4 or fewer. Likewise, preferredcycloalkyls have from 3-8 carbon atoms in their ring structure, and morepreferably have 5 or 6 carbons in the ring structure. The term C₁-C₆includes alkyl groups containing 1 to 6 carbon atoms.

[0095] Moreover, the term alkyl includes both “unsubstituted alkyls” and“substituted alkyls”, the latter of which refers to alkyl moietieshaving substituents replacing a hydrogen on one or more carbons of thehydrocarbon backbone. Such substituents can include, for example,alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Cycloalkyls can be further substituted, e.g.,with the substituents described above. An “alkylaryl” or an “arylalkyl”moiety is an alkyl substituted with an aryl (e.g., phenylmethyl(benzyl)). The term “alkyl” also includes the side chains of natural andunnatural amino acids.

[0096] The term “aryl” includes groups, including 5- and 6-memberedsingle-ring aromatic groups that may include from zero to fourheteroatoms, for example, benzene, phenyl, pyrrole, furan, thiophene,thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole,oxazole, isoxazole, pyridine, pyrazine, pyridazine, and pyrimidine, andthe like. Furthermore, the term “aryl” includes multicyclic aryl groups,e.g., tricyclic, bicyclic, e.g., naphthalene, benzoxazole,benzodioxazole, benzothiazole, benzimidazole, benzthiophene,methylenedioxyphenyl, quinoline, isoquinoline, naphthridine, indole,benzofuran, purine, benzofuran, diazapurine, or indolizine. Those arylgroups having heteroatoms in the ring structure may also be referred toas “aryl heterocycles”, “heterocycles,” “heteroaryls” or“heteroaromatics”. The aromatic ring can be substituted at one or morering positions with such substituents as described above, as forexample, halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,alkylaminoacarbonyl, arylalkyl aminocarbonyl, alkenylaminocarbonyl,alkylcarbonyl, arylcarbonyl, arylalkylcarbonyl, alkenylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety. Aryl groups can also be fused or bridged withalicyclic or heterocyclic rings which are not aromatic so as to form apolycycle (e.g., tetralin).

[0097] The term “alkenyl” includes unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double bond.

[0098] For example, the term “alkenyl” includes straight-chain alkenylgroups (e.g., ethylenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl,octenyl, nonenyl, decenyl, etc.), branched-chain alkenyl groups,cycloalkenyl (alicyclic) groups (cyclopropenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl substitutedcycloalkenyl groups, and cycloalkyl or cycloalkenyl substituted alkenylgroups. The term alkenyl further includes alkenyl groups which includeoxygen, nitrogen, sulfur or phosphorous atoms replacing one or morecarbons of the hydrocarbon backbone. In certain embodiments, a straightchain or branched chain alkenyl group has 6 or fewer carbon atoms in itsbackbone (e.g., C₂-C₆ for straight chain, C₃-C₆ for branched chain).Likewise, cycloalkenyl groups may have from 3-8 carbon atoms in theirring structure, and more preferably have 5 or 6 carbons in the ringstructure. The term C₂-C₆ includes alkenyl groups containing 2 to 6carbon atoms.

[0099] Moreover, the term alkenyl includes both “unsubstituted alkenyls”and “substituted alkenyls”, the latter of which refers to alkenylmoieties having substituents replacing a hydrogen on one or more carbonsof the hydrocarbon backbone. Such substituents can include, for example,alkyl groups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moiety.

[0100] The term “alkynyl” includes unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but which contain at least one triple bond.

[0101] For example, the term “alkynyl” includes straight-chain alkynylgroups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl,octynyl, nonynyl, decynyl, etc.), branched-chain alkynyl groups, andcycloalkyl or cycloalkenyl substituted alkynyl groups. The term alkynylfurther includes alkynyl groups which include oxygen, nitrogen, sulfuror phosphorous atoms replacing one or more carbons of the hydrocarbonbackbone. In certain embodiments, a straight chain or branched chainalkynyl group has 6 or fewer carbon atoms in its backbone (e.g., C₂-C₆for straight chain, C₃-C₆ for branched chain). The term C₂-C₆ includesalkynyl groups containing 2 to 6 carbon atoms.

[0102] Moreover, the term alkynyl includes both “unsubstituted alkynyls”and “substituted alkynyls”, the latter of which refers to alkynylmoieties having substituents replacing a hydrogen on one or more carbonsof the hydrocarbon backbone. Such substituents can include, for example,alkyl groups, alkynyl groups, halogens, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including, e.g., alkylcarbonylamino, arylcarbonylamino,carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moiety.

[0103] Unless the number of carbons is otherwise specified, “loweralkyl” as used herein means an alkyl group, as defined above, but havingfrom one to five carbon atoms in its backbone structure. “Lower alkenyl”and “lower alkynyl” have chain lengths of, for example, 2-5 carbonatoms.

[0104] The term “acyl” includes compounds and moieties which contain theacyl radical (CH₃CO—) or a carbonyl group. The term “substituted acyl”includes acyl groups where one or more of the hydrogen atoms arereplaced by for example, alkyl groups, alkynyl groups, halogens,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkyl amino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino. sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[0105] The term “acylamino” includes structures wherein an acyl moietyis bonded to an amino group. For example, the term includesalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido groups.

[0106] The term “aroyl” includes compounds and moieties with an aryl orheteroaromatic moiety bound to a carbonyl group. Examples of aroylgroups include phenylcarboxy, naphthyl carboxy, etc.

[0107] The terms “alkoxyalkyl”, “alkylaminoalkyl” and “thioalkoxyalkyl”include alkyl groups, as described above, which further include oxygen,nitrogen or sulfur atoms replacing one or more carbons of thehydrocarbon backbone, e.g., oxygen, nitrogen or sulfur atoms.

[0108] The term “alkoxy” includes substituted and unsubstituted alkyl,alkenyl, and alkynyl groups covalently linked to an oxygen atom.Examples of alkoxy groups include methoxy, ethoxy, isopropyloxy,propoxy, butoxy, and pentoxy groups. Examples of substituted alkoxygroups include halogenated alkoxy groups. The alkoxy groups can besubstituted with groups such as alkenyl, alkynyl, halogen, hydroxyl,alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkyl amino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moieties.Examples of halogen substituted alkoxy groups include, but are notlimited to, fluoromethoxy, difluoromethoxy, trifluoromethoxy,chloromethoxy, dichloromethoxy, trichloromethoxy, etc.

[0109] The term “amine” or “amino” includes compounds where a nitrogenatom is covalently bonded to at least one carbon or heteroatom. The term“alkyl amino” includes groups and compounds wherein the nitrogen isbound to at least one additional alkyl group. The term “dialkyl amino”includes groups wherein the nitrogen atom is bound to at least twoadditional alkyl groups. The term “arylamino” and “diarylamino” includegroups wherein the nitrogen is bound to at least one or two aryl groups,respectively. The term “alkylarylamino,” “alkylaminoaryl” or“arylaminoalkyl” refers to an amino group which is bound to at least onealkyl group and at least one aryl group. The term “alkylaminoalkyl”refers to an alkyl, alkenyl, or alkynyl group bound to a nitrogen atomwhich is also bound to an alkyl group.

[0110] The term “amide” or “aminocarboxy” includes compounds or moietieswhich contain a nitrogen atom which is bound to the carbon of a carbonylor a thiocarbonyl group. The term includes “alkylaminocarboxy” groupswhich include alkyl, alkenyl, or alkynyl groups bound to an amino groupbound to a carboxy group. It includes arylaminocarboxy groups whichinclude aryl or heteroaryl moieties bound to an amino group which isbound to the carbon of a carbonyl or thiocarbonyl group. The terms“alkylaminocarboxy,” “alkenylaminocarboxy,” “alkynylaminocarboxy,” and“arylaminocarboxy” include moieties wherein alkyl, alkenyl, alkynyl andaryl moieties, respectively, are bound to a nitrogen atom which is inturn bound to the carbon of a carbonyl group.

[0111] The term “carbonyl” or “carboxy” includes compounds and moietieswhich contain a carbon connected with a double bond to an oxygen atom.Examples of moieties which contain a carbonyl include aldehydes,ketones, carboxylic acids, amides, esters, anhydrides, etc.

[0112] The term “thiocarbonyl” or “thiocarboxy” includes compounds andmoieties which contain a carbon connected with a double bond to a sulfuratom.

[0113] The term “ether” includes compounds or moieties which contain anoxygen bonded to two different carbon atoms or heteroatoms. For example,the term includes “alkoxyalkyl” which refers to an alkyl, alkenyl, oralkynyl group covalently bonded to an oxygen atom which is covalentlybonded to another alkyl group.

[0114] The term “ester” includes compounds and moieties which contain acarbon or a heteroatom bound to an oxygen atom which is bonded to thecarbon of a carbonyl group. The term “ester” includes alkoxycarboxygroups such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl,butoxycarbonyl, pentoxycarbonyl, etc. The alkyl, alkenyl, or alkynylgroups are as defined above.

[0115] The term “thioether” includes compounds and moieties whichcontain a sulfur atom bonded to two different carbon or hetero atoms.Examples of thioethers include, but are not limited to alkthioalkyls,alkthioalkenyls, and alkthioalkynyls. The term “alkylthioalkyls” includecompounds with an alkyl, alkenyl, or alkynyl group bonded to a sulfuratom which is bonded to an alkyl group. Similarly, the term“alkylthioalkenyls” and alkthioalkynyls” refer to compounds or moietieswherein an alkyl, alkenyl or alkynyl group is bonded to a sulfur atomwhich is covalently bonded to an alkynyl group.

[0116] The term “hydroxy” or “hydroxyl” includes groups with an —OH or—O⁻.

[0117] The term “halogen” includes fluorine, bromine, chlorine, iodine,etc. The term “perhalogenated” generally refers to a moiety wherein allhydrogens are replaced by halogen atoms.

[0118] The terms “polycyclyl” or “polycyclic radical” refer to two ormore cyclic rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls,aryls and/or heterocyclyls) in which two or more carbons are common totwo adjoining rings, e.g., the rings are “fused rings”. Rings that arejoined through non-adjacent atoms are termed “bridged” rings. Each ofthe rings of the polycycle can be substituted with such substituents asdescribed above, as for example, halogen, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, alkoxycarbonyl, alkylaminocarbonyl,arylalkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl,arylcarbonyl, arylalkyl carbonyl, alkenylcarbonyl, aminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkyl amino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkyl, alkylaryl, or an aromatic or heteroaromatic moiety.

[0119] The term “heteroatom” includes atoms of any element other thancarbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfurand phosphorus.

[0120] It will be noted that the structure of some of the compounds ofthis invention includes asymmetric carbon atoms. It is to be understoodaccordingly that the isomers arising from such asymmetry (e.g., allenantiomers and diastereomers) are included within the scope of thisinvention, unless indicated otherwise. Such isomers can be obtained insubstantially pure form by classical separation techniques and bystereochemically controlled synthesis. Furthermore, the structures andother compounds and moieties discussed in this application also includeall tautomers thereof.

[0121] In another embodiment, the invention pertains to a method fortreating a bacterial associated state in a subject. The method includesadministering to said subject an effective amount of an oxazinonecompound of the invention, e.g., a compound of formula (I), (II), (III),(IV) or otherwise described herein.

[0122] The term “treating” includes curing as well as ameliorating atleast one symptom of the state, disease or disorder.

[0123] The term “subject” includes organisms capable of suffering froman bacterial associated state, such as mammals (e.g. primates (e.g.,monkeys, gorillas, chimpanzees, and, advantageously, humans), goats,cattle, horses, sheep, dogs, cats, mice, rabbits, pigs, dolphins,ferrets, squirrels), reptiles, or fish. In a further embodiment, thesubject is suffering from the bacterial associated disorder at the timeof administering the oxazinone compound of the invention.

[0124] The term “administering” includes routes of administration whichallow the oxazinone compound to perform its intended function. Examplesof routes of administration which can be used include parental injection(e.g., subcutaneous, intravenous, and intramuscular), intraperitonealinjection, oral, inhalation, and transdermal. The injection can be bolusinjections or can be continuous infusion. Depending on the route ofadministration, the oxazinone compound can be coated with or disposed ina selected material to protect it from natural conditions which maydetrimentally effect its ability to perform its intended function. Theoxazinone compound can be administered alone, with a pharmaceuticallyacceptable carrier, or in combination with a supplementary compound,e.g., an antibiotic, e.g., penicillin or cephalosporin. The oxazinonecompound can be administered prior to the onset of an bacterialassociated state, or after the onset of a bacterial associated state.The oxazinone compound also can be administered as a prodrug which isconverted to another (e.g., active) form in vivo.

[0125] The term “bacterial associated state” includes statescharacterized by the presence of Gram-positive pathogens, for exampleStaphylococci, Enterococci, Streptococci and mycobacteria. In a furtherembodiment, the bacterial associated state is associated with abacterial strain which is resistant to conventional antibiotics.Examples of such strains include methicillin resistant staphylococcus(MRSA), methicillin resistant coagulase negative staphylococci (MRCNS),penicillin resistant streptococcus pneumoniae and multiply resistantEnterococcus faecium.

[0126] In a further embodiment, the oxazinone compound of the inventionis administered in combination with a pharmaceutically acceptablecarrier.

[0127] In another further embodiment, the oxazinone compound isadministered in combination with a supplementary compound. Examples ofsupplementary compounds include antibiotics such a penicillin,methicillin, cephalosphorin, vancomycin, etc.

[0128] The term “in combination with” a supplementary compound isintended to include simultaneous administration of the oxazinonecompound and the supplementary compound, administration of the oxazinonecompound first, followed by the supplementary compound, andadministration of the supplementary compound first, followed by theoxazinone compound second. Any of the therapeutically useful compoundknown in the art for treating a particular bacterial associated statecan be used in the methods of the invention.

[0129] In yet another embodiment, the invention includes pharmaceuticalcompositions comprising an effective amount of a compound of theinvention, e.g., a compound of formula (I), (II), (III), (IV), orotherwise described herein.

[0130] In a further embodiment, the pharmaceutical composition of theinvention includes a pharmaceutically acceptable carrier and/or asupplementary agent, e.g., an antibiotic. In further embodiment, theeffective amount of the oxazinone compound is effective to treat abacterial infection of a subject, e.g., a human.

[0131] The invention also pertains to pharmaceutical compositionscomprising a therapeutically effective amount of a compound of theinvention and, optionally, a pharmaceutically acceptable carrier.

[0132] The language “pharmaceutically acceptable carrier” includessubstances capable of being coadministered with the compound(s) of theinvention, and which allow both to perform their intended function.Suitable pharmaceutically acceptable carriers include but are notlimited to water, salt solutions, alcohol, vegetable oils, polyethyleneglycols, gelatin, lactose, amylose, magnesium stearate, talc, silicicacid, viscous paraffin, perfume oil, fatty acid monoglycerides anddiglycerides, petroethral fatty acid esters, hydroxymethyl-cellulose,polyvinylpyrrolidone, etc. The pharmaceutical preparations can besterilized and if desired mixed with auxiliary agents, e.g., lubricants,preservatives, stabilizers, wetting agents, emulsifiers, salts forinfluencing osmotic pressure, buffers, colorings, flavorings and/oraromatic substances and the like which do not deleteriously react withthe active compounds of the invention.

[0133] The compounds of the invention that are basic in nature arecapable of forming a wide variety of salts with various inorganic andorganic acids. The acids that may be used to prepare pharmaceuticallyacceptable acid addition salts of the compounds of the invention thatare basic in nature are those that form non-toxic acid addition salts,i.e., salts containing pharmaceutically acceptable anions, such as thehydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate,phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate,citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate,succinate, maleate, gentisinate, fumarate, gluconate, glucaronate,saccharate, formate, benzoate, glutamate, methanesulfonate,ethanesulfonate, benzenesulfonate, p-toluenesulfonate and palmoate[i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)] salts. Although suchsalts must be pharmaceutically acceptable for administration to asubject, e.g., a mammal, it is often desirable in practice to initiallyisolate a compound of the invention from the reaction mixture as apharmaceutically unacceptable salt and then simply convert the latterback to the free base compound by treatment with an alkaline reagent andsubsequently convert the latter free base to a pharmaceuticallyacceptable acid addition salt. The acid addition salts of the basecompounds of this invention are readily prepared by treating the basecompound with a substantially equivalent amount of the chosen mineral ororganic acid in an aqueous solvent medium or in a suitable organicsolvent, such as methanol or ethanol. Upon careful evaporation of thesolvent, the desired solid salt is readily obtained. The preparation ofother compounds of the invention not specifically described in theforegoing experimental section can be accomplished using combinations ofthe reactions described above that will be apparent to those skilled inthe art.

[0134] The compounds of the invention that are acidic in nature arecapable of forming a wide variety of base salts. The chemical bases thatmay be used as reagents to prepare pharmaceutically acceptable basesalts of those compounds of the invention that are acidic in nature arethose that form non-toxic base salts with such compounds. Such non-toxicbase salts include, but are not limited to those derived from suchpharmaceutically acceptable cations such as alkali metal cations (e.g.,potassium and sodium) and alkaline earth metal cations (e.g., calciumand magnesium), ammonium or water-soluble amine addition salts such asN-methylglucamine-(meglumine), and the lower alkanolammonium and otherbase salts of pharmaceutically acceptable organic amines. Thepharmaceutically acceptable base addition salts of compounds of theinvention that are acidic in nature may be formed with pharmaceuticallyacceptable cations by conventional methods. Thus, these salts may bereadily prepared by treating the compound of the invention with anaqueous solution of the desired pharmaceutically acceptable cation andevaporating the resulting solution to dryness, preferably under reducedpressure. Alternatively, a lower alkyl alcohol solution of the compoundof the invention may be mixed with an alkoxide of the desired metal andthe solution subsequently evaporated to dryness.

[0135] The compounds of the invention and pharmaceutically acceptablesalts thereof can be administered via either the oral, parenteral ortopical routes. In general, these compounds are most desirablyadministered in effective dosages, depending upon the weight andcondition of the subject being treated and the particular route ofadministration chosen. Variations may occur depending upon the speciesof the subject being treated and its individual response to saidmedicament, as well as on the type of pharmaceutical formulation chosenand the time period and interval at which such administration is carriedout.

[0136] The compounds of the invention may be administered alone or incombination with pharmaceutically acceptable carriers or diluents by anyof the routes previously mentioned, and the administration may becarried out in single or multiple doses. For example, the noveltherapeutic agents of this invention can be administered advantageouslyin a wide variety of different dosage forms, i.e., they may be combinedwith various pharmaceutically acceptable inert carriers in the form oftablets, capsules, lozenges, troches, hard candies, powders, sprays,creams, salves, suppositories, jellies, gels, pastes, lotions,ointments, aqueous suspensions, injectable solutions, elixirs, syrups,and the like. Such carriers include solid diluents or fillers, sterileaqueous media and various non-toxic organic solvents, etc. Moreover,oral pharmaceutical compositions can be suitably sweetened and/orflavored. In general, the therapeutically-effective compounds of thisinvention are present in such dosage forms at concentration levelsranging from about 5.0% to about 70% by weight.

[0137] For oral administration, tablets containing various excipientssuch as microcrystalline cellulose, sodium citrate, calcium carbonate,dicalcium phosphate and glycine may be employed along with variousdisintegrants such as starch (and preferably corn, potato or tapiocastarch), alginic acid and certain complex silicates, together withgranulation binders like polyvinylpyrrolidone, sucrose, gelatin andacacia. Additionally, lubricating agents such as magnesium stearate,sodium lauryl sulfate and talc are often very useful for tablettingpurposes. Solid compositions of a similar type may also be employed asfillers in gelatin capsules; preferred materials in this connection alsoinclude lactose or milk sugar as well as high molecular weightpolyethylene glycols. When aqueous suspensions and/or elixirs aredesired for oral administration, the active ingredient may be combinedwith various sweetening or flavoring agents, coloring matter or dyes,and, if so desired, emulsifying and/or suspending agents as well,together with such diluents as water, ethanol, propylene glycol,glycerin and various like combinations thereof.

[0138] For parenteral administration (including intraperitoneal,subcutaneous, intravenous, intradermal or intramuscular injection),solutions of a therapeutic compound of the present invention in eithersesame or peanut oil or in aqueous propylene glycol may be employed. Theaqueous solutions should be suitably buffered (preferably pH greaterthan 8) if necessary and the liquid diluent first rendered isotonic.These aqueous solutions are suitable for intravenous injection purposes.The oily solutions are suitable for intraarticular, intramuscular andsubcutaneous injection purposes. The preparation of all these solutionsunder sterile conditions is readily accomplished by standardpharmaceutical techniques well known to those skilled in the art. Forparenteral application, examples of suitable preparations includesolutions, preferably oily or aqueous solutions as well as suspensions,emulsions, or implants, including suppositories. Therapeutic compoundsmay be formulated in sterile form in multiple or single dose formatssuch as being dispersed in a fluid carrier such as sterile physiologicalsaline or 5% saline dextrose solutions commonly used with injectables.

[0139] Additionally, it is also possible to administer the compounds ofthe present invention topically when treating inflammatory conditions ofthe skin. Examples of methods of topical administration includetransdermal, buccal or sublingual application. For topical applications,therapeutic compounds can be suitably admixed in a pharmacologicallyinert topical carrier such as a gel, an ointment, a lotion or a cream.Such topical carriers include water, glycerol, alcohol, propyleneglycol, fatty alcohols, triglycerides, fatty acid esters, or mineraloils. Other possible topical carriers are liquid petrolatum,isopropylpalmitate, polyethylene glycol, ethanol 95%, polyoxyethylenemonolauriate 5% in water, sodium lauryl sulfate 5% in water, and thelike. In addition, materials such as anti-oxidants, humectants,viscosity stabilizers and the like also may be added if desired.

[0140] For enteral application, particularly suitable are tablets,dragees or capsules having talc and/or carbohydrate carrier binder orthe like, the carrier preferably being lactose and/or corn starch and/orpotato starch. A syrup, elixir or the like can be used wherein asweetened vehicle is employed. Sustained release compositions can beformulated including those wherein the active component is protectedwith differentially degradable coatings, e.g., by microencapsulation,multiple coatings, etc.

[0141] In addition to treatment of human subjects, the therapeuticmethods of the invention also will have significant veterinaryapplications, e.g. for treatment of livestock such as cattle, sheep,goats, cows, swine and the like; poultry such as chickens, ducks, geese,turkeys and the like; horses; and pets such as dogs and cats. Also, thecompounds of the invention may be used to treat non-animal subjects,such as plants.

[0142] It will be appreciated that the actual preferred amounts ofactive compounds used in a given therapy will vary according to thespecific compound being utilized, the particular compositionsformulated, the mode of application, the particular site ofadministration, etc. Optimal administration rates for a given protocolof administration can be readily ascertained by those skilled in the artusing conventional dosage determination tests conducted with regard tothe foregoing guidelines.

[0143] (R)—N-Allyloxyalanine benzyl ester (2a). A solution of triflicanhydride (10.86 g, 38.5 mmol) in dry CH₂Cl₂ (20 mL) was added dropwiseto a solution of benzyl (S)-lactate (1a) (6.31 g, 35 mmol) in dry CH₂Cl₂(100 mL) with cooling (−68˜−72° C.) and stirring. After 5˜7 min, asolution of 2,6-lutidine (4.31 g, 40.3 mmol) in dry CH₂Cl₂ (15 mL) wasadded dropwise at the same temperature and the reaction mixture wasstirred for 30 min at −68˜−72° C. A solution of O-allylhydroxylamine(4.55 g, 77 mmol) in dry CH₂Cl₂ (15 mL) was added dropwise to theprepared solution of the alkylating reagent with cooling (−68˜−72° C.)and stirring. The reaction mixture was stirred for 15 min at ca. −72° C.and then was allowed to warm up to RT. After stirring for 15 h at RT,the reaction mixture was thoroughly shaken with sat. aqueous NaHCO₃ (200mL) and dried over MgSO₄. After removal of the solvent, the residue waspurified by dry flash chromatography (Silica Gel 60H for TLC, 10%EtOAc—hexanes) to afford 8.20 g (99.6%) of the product 2a as a paleyellowish oil. ¹H NMR (100 MHz) in CDCl₃+1 drop of D₂O (J, Hz): δ1.28(3H, d, ³J=7.1, MeCH), 3.82 (1H, q, ³J=7.1, MeCH), 4.22 (2H, dt, ³J=5.7,⁴J=1.2, OCH₂CH═CH₂), 5.16-5.37 (2H, m, CH═CH₂), 5.23 (2H, CH₂Ph),5.76-6.15 (1H, m, ³J=5.7, ³J^(cis)=9.9, ³J^(trans)=17.1, CH═CH₂), 7.38(5H, Ph). ¹³C (100 MHz) in CDCl₃: δ14.81, 59.09, 66.63, 75.22, 117.53,128.06, 128.24, 128.56, 134.37, 135.84, 174.01. IR (film), cm⁻: 3264,1740, 1456, 1208, 1170. Anal. Calcd for C₁₃H₁₇NO₃: C, 66.4; H, 7.3; N,5.95. Found: C, 66.3; H, 7.4; N, 6.15%.

[0144] (R)—N-(Bromoacetyl)-N-allyloxyalanine benzyl ester (3). To asolution of (R)—N-allyloxyalanine benzyl ester (2a) (2.59 g, 11 mmol)and Et₃N (1.11 g, 11 mmol) in dry CH₂Cl₂ (30 mL), a solution ofbromoacetyl bromide (2.22 g, 11 mmol) in dry CH₂Cl₂ (20 mL) was addeddropwise with cooling (−25˜−30° C.) and stirring. The reaction mixturewas stirred for 1 h at 0˜+5° C. and allowed to warm up to RT. Afterdilution with CH₂Cl₂ (ca. 30 mL), the mixture was washed with water(2×30 mL), 1N HCl (30 mL), sat. aq. NaHCO₃ (30 mL), and dried overMgSO₄. Removal of the solvent afforded 3.90 g (99%) of the product 3 asa yellowish oil. ¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ1.60 (3H, d,³J=7.3, MeCH), 4.08 (2H, CH₂Br), 4.49 (2H, dt, ³J=6.1, ⁴J=0.9,OCH₂CH═CH₂), 4.98 (1H, q, MeCH, ³J=7.3), 5.16-5.48 (2H, m, CH═CH₂), 5.20(2H, CH₂Ph), 5.76-6.15 (1H, m, ³J=6.1, ³J^(cis)=10.0, ³J^(trans)=17.3,CH═CH₂), 7.29-7.41 (5H, m, Ph). IR (film), cm⁻¹: 1746, 1678, 1455, 1214.MS (CI, isobutane), m/Z: 357 (M⁺+1), 356 (M⁺).

[0145] (R)—N-(2-triphenylphosphonio)acetyl-N-allyloxyalanine benzylester bromide or(R)—{[N-Allyloxy-N-(α-benzoxycarbonylethyl)carbamoyl]methyl}triphenylphosphoniumbromide (4). A solution of (R)—N-(Bromoacetyl)-N-allyloxyalanine benzylester (3) (3.90 g, 11 mmol) in dry toluene (10 mL) was added portionwiseto a solution of triphenylphosphine (2.885 g, 11 mmol) in dry toluene(20 mL) and the reaction mixture was stirred for 24 hours at roomtemperature. The precipitated yellow oil was separated and washed withtoluene and dried in vacuo to give a solid yellowish hygroscopic foam,which was reprecipitated from a mixture of dry CH₂Cl₂ (10 mL) andabsolute ether (ca. 100 mL) to afford 5.16 g (76%) the product 4 as awhite hygroscopic foam.

[0146] (R)—N-(2-triphenylphosphonio)acetyl-N-allyloxyalanine benzylester trifluoro methanesulfonate or(R)—{[N-Allyloxy-N-(α-benzoxycarbonylethyl)carbamoyl]methyl}triphenylphosphoniumtrifluoromethanesulfonate (5). To a solution of(R)—N-(2-triphenylphosphonio)acetyl-N-allyloxyalanine benzyl esterbromide (4) (6.18 g, 10 mmol) in dry MeCN (25 mL), a solution of silvertrifluoromethanesulfonate (2.56 g, 10 mmol) in dry MeCN (20 mL) wasadded portionwise with stirring and the yellow precipitate (AgBr) wasfiltered off using a glass-sintered funnel with a short pad of Celite545 and was washed with MeCN (ca. 50 mL). The filtrate was evaporated invacuo and the residue (yellow foam) was dissolved in CH₂Cl₂ (100 mL) andsome amount of MgSO₄ was added to the solution for coagulation of smallparticles of AgBr. The solution was filtered through a similar funnel asbefore and the filtrate was evaporated in vacuo to provide 6.52 g(99.5%) of the product 5 as a yellow highly hygroscopic foam. ¹H NMR(400 MHz) in CDCl₃ (J, Hz), major isomer: δ1.50 (3H, d, ³J=7.4, MeCH),4.84 (1H, dd, ²J=11.5, ³J=6.1, OCH_(A)CH═CH₂), 4.80 (1H, q, MeCH,³J=7.4), 4.84 (1H, dd, ²J=11.5, ³J=6.1, OCH_(B)CH═CH₂), 4.89 (1H, dd,²J_(HH)=17.8, ²J_(HP)=12.7, CH_(A)P), 5.06 (1H, d, ²J=12.4, CH_(A)Ph),5.10 (1H, d, ²J=12.4, CH_(B)Ph), 5.31 (1H, br.d, ³J^(cis)=10.4,CH═CH_(D)), 5.35 (1H, dd, ²J_(HH)=17.8, ²J_(HP)=13.8, CH_(B)P), 5.49(1H, dd, ³J^(trans)=17.2, ²J=1.2, CH═CH_(E)), 5.92-6.02 (1H, m, ³J=6.1,3J^(cis)=10.4, ³J^(trans)=17.2, CH═CH₂), 7.4-7.8 (5H, m, PhCH₂), 7.6-7.7(15H, m, Ph₃P).

[0147] (R)—N-Acryloyl-N-allyloxyalanine benzyl ester (6a). To a solutionof (R)—N-allyloxyalanine benzyl ester (2a) (7.06 g, 30 mmol) and Et₃N(4.55 g, 45 mmol) in dry CH₂Cl₂ (80 mL), a solution of acryloyl chloride(3.26 g, 36 mmol) in dry CH₂Cl₂ (20 mL) was added dropwise with cooling(−5˜−10° C.) and stirring. The reaction mixture was stirred for 1 h at0˜+5° C. and for 4.5 hours at room temperature. After dilution withCH₂Cl₂ (ca. 150 mL), the mixture was washed with water (3×50 mL), 1N HCl(50 mL), brine (100 mL), and dried over MgSO₄. Removal of the solventgave 9.16 g of a yellow oil, which was purified by dry flashchromatography (Silica Gel 60H for TLC, 3% EtOAc-hexanes→15%EtOAc-hexanes) to afford 8.52 g (98%) the product 6a as a colorless oil.¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ1.61 (3H, d, ³J=7.3, MeCH), 4.45(2H, dq, ³J=6.0, ⁴J=1.1, OCH₂CH═CH₂), 5.09 (1H, q, ³J=7.3, MeCH,), 521(2H, CH₂Ph), 5.25-5.47 (2H, m, CH═CH₂, allyl), 5.76-6.15 (1H, m,³J=6.0,³J^(cis)=10.0, ³J^(trans)=17.3, CH═CH₂, allyl), 5.83 (1H, dd, ²J=2.7,³J^(cis)=7.6, CH═CH_(A)H_(B), acryloyl), 6.48 (1H, dd, ²J=2.7,³J^(trans)=17.2, CH═CH_(A)H_(B), acryloyl), 6.80 (1H, dd, ³J^(cis)=7.6,³J^(trans)=17.2, CH═CH₂, acryloyl) 7.39 (5H, Ph). ¹³C (100 MHz) inCDCl₃: δ14.06, 56.45, 67.10, 78.27, 120.28, 126.21, 128.07, 128.21,128.48, 130.04, 130.91, 135.47, 168.22, 170.35. IR (film), cm⁻¹: 1746,1667, 1621, 1411. MS (CI, isobutane), m/Z: 290 (M⁺+1). Anal. Calcd forC₁₆H₁₉NO₄: C, 66.4; H, 6.6; N, 4.8. Found: C, 66.4; H, 6.7; N, 5.1%.

[0148] (R)—N-Acryloyl-N-allyloxyalanine tert-butyl ester (6b). Asolution of triflic anhydride (15.52 g, 55 mmol) in dry CH₂Cl₂ (25 mL)was added dropwise to a solution of tert-butyl (S)-lactate (1b) (6.71 g,50 mmol) in dry CH₂Cl₂ (130 mL) with cooling (−68˜−72° C.) and stirring.After 5˜7 min, a solution of 2,6-lutidine (6.16 g, 57.5 mmol) in dryCH₂Cl₂ (20 mL) was added dropwise at the same temperature and thereaction mixture was stirred for 1.5 h at −68˜−72° C. A solution ofO-allylhydroxylamine (6.57 g, 90 mmol) in dry CH₂Cl₂ (20 mL) was addeddropwise to the prepared solution of the alkylating reagent with cooling(−68˜−72° C.) and stirring. The reaction mixture was stirred for 15minutes at about −72° C. and then was allowed to warm up to roomtemperature. After stirring for 15 hours at room temperature, thereaction mixture was thoroughly shaken with saturated aqueous NaHCO₃(400 mL) and dried over MgSO₄. Removal of the solvent provided 14.31 gof a mixture of (R)—N-allyloxyalanine tert-butyl ester (2b) and2,6-lutidine (1:0.97 mol/mol), which was used in the next step withoutfurther purification.

[0149] To a solution of the mixture of (R)—N-allyloxyalanine tert-butylester (2b) and 2,6-lutidine in dry CH₂Cl₂ (100 mL) an additional amountof 2,6-lutidine (2.66 g, 24.8 mmol) was added and the solution wascooled to −15˜−20° C. A solution of acryloyl chloride (5.09 g, 56.3mmol) in dry CH₂Cl₂ (20 mL) was added dropwise at the same temperature.The reaction mixture was stirred for 1 h at 0˜+5° C. and for 20 hours atroom temperature. After dilution with CH₂Cl₂ (about 150 mL), the mixturewas washed with water (4×50 mL), 1M citric acid (2×50 mL), saturatedaqueous NaHCO₃ (2×50 mL) and dried over MgSO₄. Removal of the solventgave 12.73 g of a red oil, which was purified by dry flashchromatography (Silica Gel 60H for TLC, 1% EtOAc-hexanes→7%EtOAc-hexanes) to afford 8.91 g (70%) of the product 6b as a colorlessoil. ¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ1.47 (9H, Me₃C), 1.53 (3H, d,³J=7.3, MeCH), 4.48 (2H, dq, ³J=5.9, ⁴J=1.2, OCH₂CH═CH₂), 4.93 (1H, q,³J=7.3, MeCH,), 5.26-5.50 (2H, m, CH═CH₂, allyl), 5.76-6.18 (1H, m,³J=5.9, ³J^(cis)=10.1, ³J^(trans)=15.6, CH═CH₂, allyl), 5.81 (1H, dd,²J=2.6, ³J^(cis)=9.5, CH═CH_(A)H_(B), acryloyl), 6.46 (1H, dd, ²J=2.6,³J^(trans)=17.0, CH═CH_(A)H_(B), acryloyl), 6.80 (1H, dd, ³J^(cis)=9.5,³J^(trans)=17.0, CH═CH₂, acryloyl). ¹³C (100 MHz) in CDCl₃: δ14.12,27.91, 57.28, 78.22, 81.83, 120.02, 126.42, 129.75, 131.09, 132.41,168.34, 169.55. IR (film), cm⁻¹: 2981, 1738, 1667, 1622, 1411. MS (CI,isobutane), m/Z: 256 (M⁺+1).

[0150] (R)-2-(α-Benzoxycarbonylethyl)-4,5-dehydro-1,2-oxazinan-3-one or(R)-2-(3-Oxo-3,6-dihydro-[1,2]oxazin-2-yl)-propionic acid benzyl ester(7a).

[0151] Method A. A solution of(R)—N-(2-triphenylphosphonio)acetyl-N-allyloxyalanine benzyl estertrifluoromethanesulfonate (5) (6.52 g, 9.95 mmol) in CH₂Cl₂ (180 mL) wascooled to −76˜−78° C. and saturated with ozone (a moderate bubblinguntil a persistent blue color) at the same temperature with stirring. Anexcess of ozone was removed by bubbling N₂ through the reaction mixtureat −76˜−78° C. and Me₂S (3.1 g, 50 mmol) was added portionwise withstirring at the same temperature. The reaction mixture was allowed towarm up to room temperature and a solution of K₂CO₃ (6.91 g, 50 mmol) inwater (20 mL) was added. After a vigorous stirring, the reaction mixturewas diluted with water (100 mL) and the organic layer was separated offand washed with water (2×30 mL) and dried over MgSO₄. Removal of thesolvent gave a yellow semi-solid substance, which was purified by flashchromatography (Silica Gel 60, 20% EtOAc-hexanes) to afford 0.70 g (27%) the product 7a as a colorless oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz):δ1.53 (3H, d, ³J=7.3, MeCH), 4.46 (1H, dq, ²J=−15.9, ³J=3.4, ⁴J=1.8,OCH_(A)CH═CH), 4.56 (1H, dq, ²J=15.9, ³J=3.4, ⁴J=1.8, OCH_(B)CH═CH),5.15 (1H, d, ²J=12.4, CH_(A)Ph), 5.19 (1H, q, MeCH, ³J=7.3), 5.22 (1H,d, ²J=12.4, CH_(B)Ph), 6.04 (1H, dt, ³J^(cis)=10.0, ⁴J=1.8, CH═CHCO),6.72 (1H, dt, ³J^(cis)=10.0, ³J=3.4, CH₂CH═CHCO), 7.27-7.40 (5H, m, Ph).¹³C (100 MHz) in CDCl₃: δ13.71, 53.57, 67.08, 67.57, 122.32, 128.02,128.26, 128.52, 135.49, 139.67, 164.64, 170.09. IR (film), cm⁻¹: 1745,1675, 1214, 1189. MS (CI, isobutane), m/Z: 262 (M⁺+1). Anal. Calcd forC₁₄H₁₅NO₄: C, 64.4; H, 5.8; N, 5.4. Found: C, 64.3; H, 5.85; N, 5.6%.

[0152] Method B. A solution of(R)-N-acryloyl-N-allyloxyalanine benzylester (6a) (7.32 g, 25.3 mmol) andbis(tricyclohexylphosphine)benzylidene ruthenium dichloride (Grubb'scatalyst) (1.04 g, 1.26 mmol, 5 mol %) in dry CH₂Cl₂ (250 mL) wasrefluxed with Silica Gel 60H (for TLC) followed by washing with CH₂Cl₂(50 mL). After removal of the solvent, the dark oily residue waspurified by dry flash chromatography (Silica Gel 60H for TLC, 5%EtOAc-hexanes→15% EtOAc-hexanes) to afford 6.05 g (92%) the olefin 7aidentical to that described above.

[0153] (R)-2-(α-tert-Butoxycarbonylethyl)-4,5-dehydro-1,2-oxazinan-3-oneor (R)-2-(3-Oxo-3,6-dihydro-[1,2]oxazin-2-yl)-propionic acid tert-butylester (7b). A solution of (R)-N-acryloyl-N-allyloxyalanine tert-butylester (6b) (3.39 g, 13.27 mmol) andbis(tricyclohexylphosphine)benzylidene ruthenium dichloride (Grubb'scatalyst) (0.481 g, 0.584 mmol, 4.4 mol %) in dry benzene (120 mL) wasrefluxed with stirring for 6 hours in N₂ atmosphere. Two portions (0.234g, 0.284 mmol and 0.209 g, 0.254 mmol) of the catalyst were addedsuccessively and the reaction mixture was refluxed for 8 hours aftereach addition. The dark brown reaction mixture was filtered through ashort pad of Silica Gel 60H (for TLC) followed by washing with CH₂Cl₂(100 mL). After removal of the solvent, the dark oily residue waspurified by dry flash chromatography (Silica Gel 60H for TLC, 5%EtOAc-hexanes→20% EtOAc-hexanes) to afford 2.45 g (81%) the olefin 7b asa colored oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ1.46 (9H, Me₃C), 1.53(3H, d, 3J=7.3, MeCH), 4.49 (1H, dq, ²J=15.8, ³J=3.4, ⁴J=1.8,OCH_(A)CH═CH), 4.64 (1H, dq, ²J=−15.8, ³J=3.4, ⁴J=1.8, OCH_(B)CH═CH),5.03 (1H, q, MeCH, ³J=7.3), 6.05 (1H, dt, ³J^(cis)=10.0, ⁴J=1.8,CH═CHCO), 6.74 (1H, dt, ³J^(cis)=10.0, ³J=3.4, CH₂CH═CHCO). ¹³C (100MHz) in CDCl₃: δ13.77, 27.95, 54.16, 67.53, 81.82, 122.46, 139.47,164.67, 169.28. IR (film), cm⁻¹: 2979, 1737, 1677, 1368, 1162.

[0154] MS (CI, isobutane), m/Z: 228 (M⁺+1). Anal. Calcd for C₁₁H₁₇NO₄:C, 58.1; H, 7.5; N, 6.2. Found: C, 58.4; H, 7.7; N, 6.3%.

[0155] N-Allyloxyphthalimide. To a cooled solution (5 to 10° C.) ofN-hydroxyphthalimide (53.80 g, 0.33 mol) in DMF (120 mL), was added DBU(46.36 mL, 0.31 mol) was added in one portion, with stirring. After 10minutes, allyl bromide (25.96 mL, 0.30 mol) was added in portions. Thecooling bath was removed and the reaction mixture was stirred at roomtemperature for 1 hour, until the color turned from dark burgundy toyellow. Ethyl acetate (300 mL) and water (150 mL) were added, theaqueous phase was separated, and the organic phase was washed with water(4×50 mL). The combined aqueous extracts were back-extracted with EtOAc(3×50 mL) and the total EtOAc solution was washed with saturated NaHCO₃(10×50 mL) until the aqueous phase was pale yellow, then with brine(2×75 mL) and dried over anhydrous MgSO₄. Removal of the solventafforded 58.83 g (96% yield) of the product as a white solid.

[0156] Allyloxyamine. N-Allyloxyphthalimide (58.70 g, 0.289 mol) wasdissolved in absolute EtOH (295 mL) with heating and stirring. Then, asolution of N₂H₄.H₂O (14.01 mL, 0.289 mol) in EtOH (44 mL) was addedportionwise, with stirring. The reaction mixture immediately turnedyellow and a white precipitate formed. The mixture was refluxed for 2hours, stirred at room temperature for 17 hours, cooled (10 to 15° C.),and concentrated HCl (30.24 mL, 0.369 mol) was added portionwise. Thewhite precipitate was removed by filtration and washed with water (160mL). The combined filtrates were evaporated under reduced pressure andthe residue was maintained at 1 torr for 3 hours. Water (100 mL) wasadded, the solution was filtered, the filtrate was concentrated to 50 mLand added portionwise to solid KOH pellets in a distillation apparatus.The product was collected at 80-100° C. A second distillation over KOHpellets afforded 18.958 g (89% yield) of the product as a colorlessliquid, b.p. 94-97° C.

[0157] (S)-2-Hydroxy-3-methylbutyric acid benzhydryl ester (2f). Asolution of diphenyldiazomethane (0.70 g, 3.604 mmol) in EtOAc (5 mL)was added portionwise with cooling (ice bath) and stirring to a solutionof (S)-2-hydroxy-3-methylbutyric acid (0.51 g, 4.355 mmol) in EtOAc (5mL). The resulting purple solution was stirred at room temperature for 6h until color faded to pale yellow, and then was washed successivelywith saturated aqueous NaHCO₃ (3×5 mL) and brine (2×5 ML), dried overanhydrous MgSO₄ and evaporated to give 0.9864 g of a yellow solid.Recrystallization from hexanes yielded 0.82 g (80%) of 2f as soft whitecrystals (m.p. 71-72° C.). ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ0.7638(d, 3H, —CH₃), 1.0181 (d, 3H, —CH₃), 2.1229-2.2346 (m, 1H, CHMe₂),2.6768 (br. s, 1H, OH), 4.1612 (d, 1H, CHOH), 6.96 (s, 1H, CHPh₂),7.20-7.45 (m, 10H, ArH). MS (CI, isobutane), m/z: 283 (M⁺). Anal. Calcd.for C₁₈H₂₀O₃: C, 76.03; H, 7.09. Found: C, 76.13; H, 7.12%.

[0158] (S)-2-Trifluoromethanesulfonyloxy-3-methylbutyric acid benzhydrylester. A solution of triflic anhydride (4.42 mL, 26.29 mmol) in dryCH₂Cl₂ (12 mL) was added dropwise via syringe to a solution of 2f (6.50g, 22.86 mmol) in dry CH₂Cl₂ (70 mL), with cooling (−68 to −70° C.) andstirring. After 5 min, a solution of 2,6-lutidine (3.19 mL, 27.43 mmol)in dry CH₂Cl₂ (10 mL) was added dropwise. The pale yellow reactionmixture was stirred at −68° C. for 1 h, warmed to room temperature andstirred for an additional 17 h. The solvent was removed under reducedpressure and the residue (in EtOAc) was washed with 1N citric acid(5×100 mL), dried over anhydrous MgSO₄ and evaporated to give 9 g of ared semi-solid. This was purified via dry flash chromatography (SilicaGel 60H for TLC, 5% EtOAc-hexanes) to afford 8.04 g of the triflate as apale yellow solid. ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ0.8878 (d, 3H,—CH₃), 1.0479 (d, 3H, —CH₃), 2.3842-2.4628 (m, 1H, CHMe₂), 5.0431 (d,1H, CHOSO₂CF₃), 7.000 (s, 1H, CHPh₂), 7.20-7.42 (m, 10H, ArH). MS (CI,isobutane), m/z: 415 (M⁺−1). Anal. Calcd. for C₁₉H₁₉O₅SF₃: C, 54.80; H,4.60. Found: C, 54.55; H, 4.63%.

[0159] (R)-2-Allyloxyamino-3-methylbutryic acid benzhydryl ester (3f). Asolution of O-allyloxylamine (5.414 g, 0.0741 mol) in dry CH₃CN (35 mL)was added, with stirring, to a solution of the triflate (7.711 g, 0.0185mol) in dry CH₃CN (80 mL). The reaction mixture was refluxed for 3 h,cooled to room temperature, and evaporated under reduced pressure. Theresidue, in EtOAc (100 mL), was washed successively with saturatedNaHCO₃ (3×100 mL) and brine (2×100 mL), dried over anhydrous MgSO₄ andevaporated to give 6.6720 g of a pale yellow oil. Purification by dryflash chromatography (Silica Gel 60H for TLC, 10% EtOAc-hexanes)afforded 5.6229 g (89%) of 3f as a colorless oil. ¹H NMR (400 MHz) inCDCl₃ (J, Hz): δ0.8650 (d, 3H, —CH₃), 0.9150 (d, 3H, —CH₃),1.8524-1.9379 (m, 1H, CHMe₂),3.5712 (d, 1H, NCH), 4.1667-4.1929 (m, 2H,OCH₂), 5.1422-5.1780 (dm, 1H, OCH₂CH═CHH), 5.2223 (dd, 1H, OCH₂CH═CHH),5.8281-5.9268 (m, 1H, OCH₂CH), 6.975 (s, 1H, CHPh₂), 7.23-7.40 (m, 10H,ArH). MS (CI, isobutane), m/z: 340 (M⁺+1). Anal. Calcd. for C₂₁H₂₅NO₃:C, 74.31; H, 7.42; N, 4.13. Found: C, 74.49; H, 7.46; N, 4.40%.

[0160] (R)-2-(N-Acryloyl, N-allyloxyamino)-3-methylbutyric acidbenzhydryl ester (6f). A solution of 3f (5.085 g, 14.981 mmol) in dryCH₂Cl₂ (50 mL) was treated in one portion with triethylamine (3.13 mL,22.471 mmol. The solution was stirred at room temperature for 5 min,cooled to 0° C. and 97% acryloyl chloride (1.50 mL, 17.977 mmol) wasadded dropwise via syringe. This mixture was stirred at 0° C. for 45min, then allowed to warm to room temperature, and stirring wascontinued for 2.5 h. The light orange reaction mixture was then dilutedwith CH₂Cl₂ (150 mL) and washed successively with water (3×50 mL), N HCl(50 mL) and brine (100 mL), dried over anhydrous MgSO₄ and evaporated to5.6721 g of an orange oil. Purification by dry flash chromatography(Silica Gel 60H for TLC, 3% EtOAc-hexanes→5% EtOAc-hexanes) gave 4.7904g (81%) of 6f as a yellow oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz):δ0.9465 (d, 3H, —CH₃), 0.9631 (d, 3H, —CH₃), 2.4434-2.5352 (m, 1H,CHMe₂), 4.1484 (dd, 1H, OCHH), 4.3206 (dd, 1H, OCHH), 4.8691 (d, 1H,NCH), 5.1729-5.2203 (m, 2H, OCH₂CH═CH₂), 5.7255-5.8248 (m, 2H,OCH₂CH+NC═OCH), 6.4773 (dd, 1H, NC═OCH═CHH), 6.7104 (dd, 1H,NC═OCH═CHH), 6.9054 (s, 1H, CHPh₂), 7.20-7.40 (m, 10H, ArH). MS (CI,isobutane), m/z: 397 (M⁺+1).

[0161] (R)-3-Methyl-2-(3-oxo-3,6-dihydro-[1,2]oxazin-2-yl)-butyric acidbenzhydryl ester (7f). Bis(tricyclohexylphosphine)benzylidene rutheniumdichloride (0.343 g, 0.417 mmol) was weighed directly into a 250 mLflask, placed under a N₂ atmosphere, and dry CH₂Cl₂ (25 mL) was added,followed by a solution of 6f (3.279 g, 8.333 mmol) in dry CH₂Cl₂ (60mL). The dark purple reaction mixture was refluxed for 4 h, cooled toroom temperature, filtered through a short, densely-packed pad of SilicaGel 60H for thin layer chromatography (φ˜3.8 cm, 1˜2 cm), which waswashed with CH₂Cl₂ (5×20 mL). Removal of the solvent gave 3.175 g of abrown oil which was purified via dry flash chromatography (Silica Gel60H for TLC, 5% EtOAc-hexanes→20% EtOAc-hexanes) to afford 3.5682 g(90%) of 7f as a brown oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ0.9897(d, 3H, —CH₃), 1.0067 (d, 3H, —CH₃), 2.4435-2.5324 (m, 1H, CHMe₂),4.3791 (ddd, 2H, NOCH₂), 4.9721 (d, 1H, NCH), 6.0472 (dt, 1H,NOCH₂CH═CH), 6.6793 (dt, 1H, NOCH₂CH), 6.9187 (s, 1H, CHPh₂), 7.20-7.40(m, 10H, ArH). MS (CI, isobutane), m/z: 366 (M⁺+1).

[0162] (R)-3-Methyl-2-(3-oxo-3,6-dihydro-[1,2]oxazin-2-yl)-butyric acid(8f, R═H). A solution of 7d (2.00 g, 5.473 mmol) in 98% formic acid (120mL) was stirred vigorously at room temperature for 6 h. The solvent wasthen removed in vacuo, and the residue, in dry CH₂Cl₂ (100 mL), wasdried over anhydrous MgSO₄. Removal of the solvent gave 2.2220 g of ayellow oil which was purified via dry flash chromatography (Silica Gel60H for TLC, 30% EtOAc-hexanes→100% EtOAc) to afford 0.8185 g (75%) of8f as a yellow solid. ¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ1.10 (d, 3H,—CH₃), 1.15 (d, 3H, —CH₃), 2.20-2.80 (m, 1H, CHMe₂), 4.40-4.75 (m, 2H,NOCH₂), 4.90 (d, 1H, NCH), 6.15 (dt, 1H, NOCH₂CH═CH), 6.81 (dt, 1H,NOCH₂CH), 8.33 (br. s, 1H, CO₂H). MS (CI, isobutane), m/z: 200 (M⁺).

[0163] (R)-3-Methyl-2-(3-oxo-[1,2]oxazinan-2-yl)-butyric acid (9f) Asuspension of 10% Pd on activated carbon (0.032 g) in EtOAc (5 mL) wasstirred for 1 h in a H₂ atmosphere and a solution of 8f (0.032 g, 0.1606mmol) was added in one portion. This reaction mixture was stirred for 24hours at room temperature, and then filtered through a densely-packedpad of Silica Gel 60H for thin layer chromatography, which was washedwith EtOAc (4×5 mL). Removal of the solvent afforded 0.0193 g (60%) of9f as a white solid. ¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ1.02 (d, 3H,—CH₃), 1.11 (d, 3H, —CH₃), 1.98-2.35 (m, 2H, NOCH₂CH₂), 2.35-2.80 (m,3H, NOCH₂CH₂CH₂+CHMe₂), 4.00-4.45 (m, 2H, NOCH₂), 4.85 (d, 1H, NCH),6.20 (br. s, 1H, CO₂H). MS (CI, isobutane), m/z: 202 (M⁺).

[0164] (S)-Hydroxy-phenylacetic acid benzhydryl ester (2g). A solutionof diphenyldiazomethane (0.528 g, 2.719 mmol) in EtOAc (5 mL) was addedportionwise with cooling (ice bath) and stirring to a solution of(S)-mandelic acid (0.500 g, 3.286 mmol) in EtOAc (5 mL). The purplereaction mixture was stirred at room temperature for 1 h and was thenwashed successively with saturated NaHCO₃ (3×5 mL) and brine (2×5 mL),dried over anhydrous MgSO₄ and evaporated to 0.9864 g of a yellow solid,which was recrystallized from hexanes to give 0.67 g (80%) of 2g. ¹H NMR(400 MHz) in CDCl₃ (J, Hz): δ3.4515 (d, 1H, OH), 5.2782 (d, 1H, CHOH),6.75 (s, 1H, CHPh₂), 7.20-7.45 (m, 15H, ArH).

[0165] (R)-Allyloxyamino phenylacetic acid benzhydryl ester (3g). Asolution of triflic anyhydride (10.80 mL, 64.471 mmol) in dry CH₂Cl₂ (25mL) was added dropwise via syringe to a solution of 2g (17.85 g, 56.062mmol) in dry CH₂Cl₂ (175 mL), with cooling (−68 to −70° C.) andstirring. After 5 min, a solution of 2,6-lutidine (7.80 mL, 67.275 mmol)in dry CH₂Cl₂ (25 mL) was added dropwise. The pale yellow reactionmixture was stirred at −68° C. for 1 h, and a solution of allyloxyamine(7.294 g, 99.791 mmol) in dry CH₂Cl₂ (25 mL) was then added dropwisewith continued cooling and stirring. The yellow reaction mixture wasstirred for 30 minutes and the cooling bath was then removed and themixture allowed to warm to room temperature. Stirring was continued atroom temperature for 17 hours and the dark yellow solution was mixed ina separatory funnel with 250 mL of saturated NaHCO₃. The two layers weregently mixed by bubbling N₂ for 20 min, and then by thorough manualshaking. The organic phase was separated, washed successively withsaturated NaHCO₃ (2×250 mL) and brine (250 mL), dried over anhydrousMgSO₄ and evaporated to 24.048 g of a mixture of 3g and 2,6-lutidine(1:0.37 mol:mol), which was used in the next step without furtherpurification. ¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ4.28 (dt, 2H, NOCH₂),4.95 (br. d, 1H, NCH), 5.10-5.45 (m, 2H, NOCH₂CH═CH₂), 5.72-6.15 (m, 1H,NOCH₂CH), 6.22 (br. d, 1H, NH), 6.98 (s, 1H, CHPh₂), 7.02-7.50 (m, 10H,ArH).

[0166] (R)-(N-Acryloyl, N-allyloxyamino)-phenylacetic acid benzhydrylester (6g). 2,6-Lutidine (4.24 mL, 36.3972 mmol) was added dropwise,with stirring, to a solution of 3g (9.0617 g, 24.2648 mmol) in dryCH₂Cl₂ (75 mL dry CH₂Cl₂ (25mL). After 5 minutes at room temperature,the reaction mixture was cooled to 0° C. and a solution of 97% acryloylchloride (2.44 mL, 29.1178 mmol) in dry CH₂Cl₂ (25 mL) was addedportionwise with stirring. This mixture was stirred at 0° C. for 45minutes, then allowed to warm to room temperature and stirred for 17hours. The reaction mixture was then diluted with CH₂Cl₂ (50 mL) andwashed successively with water (50 mL), N citric acid (2×100 mL),saturated NaHCO₃ (2×75 mL) and brine (100 mL), dried over anhydrousMgSO₄ and evaporated to a dark orange oil which was purified via dryflash chromatography (Silica Gel 60H for TLC, 3% EtOAc-hexanes→20%EtOAc-hexanes) to afford 6.50 g (32%) of 7g as a yellow oil. ¹H NMR (400MHz) in CDCl₃ (J, Hz): δ3.5989 (dd, 1H, NOCHH), 4.0682 (dd, 1H, NOCHH),5.0355 (dd, 1H, NOCH₂CH═CHH), 5.1036 (dd, 1H, NOCH₂CH═CHH),5.5341-5.6340 (m, 1H, NOCH₂CH), 5.8435 (dd, 1H, C═OCH), 6.2839 (s, 1H,NCH), 6.4936 (dd, C═OCHCHH), 6.7685 (dd, 1H, C═OCHCHH), 6.9356 (s, 1H,CHPh₂), 7.02-7.46 (m, 15H, ArH). MS (CI, isobutane), m/z: 428 (M⁺+1).

[0167] (R)-(3-Oxo-3,6-dihydro-[1,2]oxazin-2-yl)-phenylacetic acidbenzhydryl ester (7g). Bis(tricyclohexylphosphine)benzylidene rutheniumdichloride (0.11 g, 0.1289 mmol) was weighed directly into a 50 mLflask, placed under a N₂ atmosphere, and dry CH₂Cl₂ (10 mL) was added,followed by a solution of 6g (1.1020 g, 2.5778 mmol) in dry CH₂Cl₂ (15mL). The purple reaction mixture was refluxed for 3 h, an additional0.11 of catalyst was added, and refluxing was continued for another 2hours. The mixture was then allowed to cool to room temperature andfiltered through a short, densely-packed pad of Silica Gel 60H for thinlayer chromatography (φ˜3.8 cm, 1˜2 cm), which was washed with CH₂Cl₂(5×20 mL). Evaporation gave a brown oil which was purified via columnchromatography (5% EtOAc-hexanes→30% EtOAc-hexanes) to afford 0.500 g(50% yield) of 7g as a brown oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz):δ4.3375 (ddd, 1H, NOCHH), 4.6139 (ddd, 1H, NOCHH), 6.0447 (dt, 1H,NOCH₂CH═CH), 6.3715 (s, 1H, NCH), 6.7020 (dt, 1H, NOCH₂CH), 6.9626 (s,1H, CHPh₂), 7.10-7.40 (m, 15H, ArH). MS (CI, isobutane), m/z: 400(M⁺+1).

[0168] (R)-(3-Oxo-3,6-dihydro-[1,2]oxazin-2-yl)-phenylacetic acid (8g).A solution of 7e (0.150 g, 0.3755 mmol) in 98% formic acid (10 mL) wasstirred vigorously at room temperature for 6 hours. The solvent was thenremoved, and any residual acid was co-evaporated with dry Et₂O (2×5 mL).The residue was dissolved in dry CH₂Cl₂ (15 mL), dried over anhydrousMgSO₄ and evaporated to a yellow oil, which was purified via columnchromatography (40%EtOAc-hexanes→100% EtOAc) to afford 0.070 g (80%) of8g as a brown solid. ¹H NMR (100 MHz) in CDCl₃ (J, Hz): δ4.41 (ddd, 1H,NOCHH), 4.78 (ddd, 1H, NOCHH), 6.12 (dt, 1H, NOCH₂CH═CH), 6.37 (s, 1H,NCH), 6.78 (dt, 1H, NOCH₂CH), 7.15-7.60 (m, 5H, ArH), 8.40-9.10 (br. s,1H, CO₂H).

[0169](αR,4S,5S)-2-(α-Benzoxycarbonylethyl)-4,5-dihydroxy-1,2-oxazinan-3-one(8a). A mixture of K₂OsO₂(OH)₄ (0.0063 g, 0.017 mmol), (DHQD)₂PHAL(0.032 g, 0.041 mmol), and AD-mix-β (1.41 g) was dissolved in a mixtureof tert-butanol (5 mL) and water (5 mL). After dissolution of the salts,methanesulfonamide (0.0994 g, 1.04 mmol) was added and the mixture wascooled to −2˜0° C.(R)-2-(α-Benzoxycarbonylethyl)-4,5-dehydro-1,2-oxazinan-3-one (7a)(0.264 g, 1.01 mmol) was added in one portion and the heterogeneousslurry was stirred vigorously for 14 h at −2˜+2° C. Sodium sulfite (1.5g) was added in order to quench the oxidation and stirring was continuedfor 1 h at RT. CH₂Cl₂ (10 mL) was added and the upper organic layer wasseparated off. The aqueous phase was extracted with CH₂Cl₂ (5×5 mL). Thecombined organic extracts were dried over MgSO₄. After removal of thesolvent, the yellowish semisolid residue was purified by dry flashchromatography (Silica Gel 60H for TLC, 15% EtOAc-hexanes→EtOAc) toafford 0.158 g (81%) the product 8a as a white solid. ¹H NMR (400 MHz)in CDCl₃+2 drops of CD₃OD (J, Hz): δ1.53 (3H, d, ³J=7.3, α-MeCH), 3.96(1H, dd, ²J=−11.8, ³J=2.5, 6-H_(A)), 4.41 (1H, d, ³J=4.1, 4-H), 4.44(1H, dd, ²J=−11.8, ³J=6.8, 6-H_(B)), 4.55−4.51 (1H, m, 5-H), 5.11 (1H,q, ³J=7.3, α-CH), 5.13 (1H, s, CH₂Ph), 7.34−7.24 (5H, m, Ph).

[0170](αR,4R,5R)-2-(α-tert-Butoxycarbonylethyl)-4,5-dihydroxy-1,2-oxazinan-3-one(8b) was obtained as a white solid (mp 93-94° C.) with a yield of 83%from (R)-2-(α-tert-butoxycarbonylethyl)-4,5-dehydro-1,2-oxazinan-3-one(7b) similarly to the preparation of 8a (vide infra) using (DHQ)₂PHAL asthe asymmetric catalyst. ¹H NMR (400 MHz) in CDCl₃+2 drops of CD₃OD (J,Hz): δ1.44 (3H, d, ³J=7.3, α-MeCH), 1.45 (9H, Me₃C), 2.25 (2H, br. s,OH, OH), 4.30 (1H, dd, ²J=−11.9, ³J=2.8, 6-H_(A)), 4.37 (1H, d, ³J=4.5,4-H), 4.38 (1H, dd, ²J=−11.9,³J=7.3, 6-H_(B)), 4.56−4.52 (1H, br. m,5-H), 4.96 (1H, q, ³J=7.3, α-CH). ¹³C (100 MHz) in CDCl₃: δ13.96, 27.92,54.89, 68.89, 69.98, 76.55, 83.01, 169.46, 169.94. IR (KBr), cm⁻¹: 3423,2985, 1739, 1662, 1415, 1152. MS (CI, isobutane), m/Z: 261 (M⁺+1). Anal.Calcd for C₁₁H₁₉NO₆: C, 50.6; H, 7.3; N, 5.4.

[0171] Found: C, 50.5; H, 7.4; N, 5.4%.

[0172] (αR,4S,5S)-2-(α-Carboxyethyl)-4,5-dihydroxy-1,2-oxazinan-3-one(9a). A suspension of 5% Pd-C (0.137 g) in EtOAc (10 mL) was stirred for1 h under H₂ atmosphere and a solution of the benzyl ester 8a (0.137 g,0.464 mmol) in EtOAc (15 ml) was added portionwise. The reaction mixturewas stirred for 8 hours under H₂ and filtered through a short pad ofSilica Gel 60H (for TLC) followed by washing with MeOH (7×5 mL). Afterevaporation of the filtrate in vacuo, the residue was triturated withdry ether and dried in vacuo to provide 0.093 g (98%) of the product 9aas a white solid. ¹H NMR (400 MHz) in CD₃CN (J, Hz): δ1.44 (3H, d,³J=7.3, α-MeCH), 3.00 (2H, br. s, OH, OH), 3.90 (1H, m, 6-H_(A)), 4.37(1H, d, ³J=4.4, 4-H), 4.49-4.43 (2H, m, 6-H_(B), 5-H), 5.01 (1H,q,³J=7.3, α-CH), 5.31 (1H, br. s, COOH). ¹³C (100 MHz) in CD₃CN: δ13.55,54.34, 70.15, 70.64, 78.41, 171.22, 172.85.

[0173] (αR,4R,5R)-2-(α-Carboxyethyl)-4,5-dihydroxy-1,2-oxazinan-3-one(9b). To a cooled (ca. 0° C.) stirred solution of the tert-butyl ester8b (0.0467 g, 0.179 mmol) in CH₂Cl₂ (0.9 ml), trifluoroacetic acid (0.9mL) was added in one portion. The reaction mixture was kept for 1 hourat room temperature and was evaporated in vacuo to dryness. Thesemisolid residue was triturated with dry ether and dried in vacuo toafford 0.0295 g (80%) of the product 9b as a white solid. ¹H NMR (400MHz) in CD₃CN (J, Hz): δ1.41 (3H, d, ³J=7.3, α-MeCH), 3.60 (2H, br. s,OH, OH, COOH), 4.14 (1H, dd, ²J=−11.6,³J=2.3, 6-H_(A)), 4.34 (1H, d,³J=4.1, 4-H), 4.40 (1H, dd, ²J=−11.6, ³J=7.4, 6-H_(B)), 4.49−4.45 (2H,m, 5-H), 5.01 (1H, q, ³J=7.3, α-CH), 5.31 (1H, br. s, COOH). ¹³C (100MHz) in CD₃CN: δ14.45, 54.48, 70.05, 70.99, 77.77, 167.30, 171.82.

[0174](αR,1R,6R)-2-(α-tert-Butoxycarbonylethyl)-8-thia-3,7,9-trioxa-4-azabicyclo[4.3.0]nona-5,8-dione(10). To a solution of the diol 8b (2.395 g, 9.167 mmol) and Et₃N (3.71g, 36.67 mmol) in dry CH₂Cl₂ (45 mL), a solution of thionyl chloride(1.64 g, 13.75 mmol) in dry CH₂Cl₂ (15 mL) was added dropwise withcooling (−15˜−20° C.) and stirring. After stirring for 1 hour at −2˜2°C., the reaction mixture was diluted with CH₂Cl₂ (60 mL) and washed withwater (20 mL), 1M aqueous citric acid (20 mL), saturated aqueous NaHCO₃(20 mL) and dried over MgSO₄. Removal of the solvent provided 2.524 g(90%) of the product 10 (diastereomer ratio is ca. 1:1) as a viscousdark, oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ1.45 (9H, Me₃C), 1.46(9H, Me′₃C), 1.47 (3H, d, ³J=7.3, α-MeCH), 1.49 (3H, d,³J=7.3, α-Me′CH),4.43 (1H, dd, ²J=−12.4, ³J=5.5,2-H_(A)), 4.48(1H, dd, ²J=−11.5, ³J=5.0,²-H_(A)′), 4.51 (1H, dd, ²J=−11.5, ³J=5.0, ²-H_(B)), 4.60 (1H, dd,²J=−12.4, ³J=6.5, ²-H_(B)′), 5.01 (1H, q, ³J=7.3, α-CH), 5.05 (1H, q,α-CH, ³J=7.3), 5.18 (1H, d, ³J=8.2, 6-H), 5.22 (1H, m, ³J=8.2, 6.5, 5.5,1-H), 5.37 (1H, d, ³J=7.3, 6-H′), 5.50 (1H, m, ³J=7.3, 5.0, 1-H′). Thecompound was used in the next reaction without further purification.

[0175](αR,4S,5R)-2-(α-tert-Butoxycarbonylethyl)-4-azido-5-hydroxy-1,2-oxazinan-3-one(11) To a stirred solution of the cyclic sulfite (2,524 g, 8.21 mmol) inHMPA (15 mL), sodium azide (1.60 g, 24.6 mmol) was added in one portionand the reaction mixture was stirred for 24 hours at room temperature.EtOAc (50 mL) was added and the resultant mixture was washed with water(2×15 ml,) and 1M aqueous citric acid (15 mL). The combined aqueoussolutions were extracted with EtOAc (3×15 mL). The combined organicextracts were washed with saturated aqueous NaHCO₃ (50 mL), brine (2×50mL) and dried over MgSO₄. After removal of the solvent, the dark oilyresidue was purified by flash chromatography (Silica Gel 60, 1%MeCN—CH₂Cl₂→10% MeCN—CH₂Cl₂) to afford 0.37 g (16 %) the product 11 as ayellowish oil. ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ1.46 (9H, Me₃C), 1.48(3H, d, ³J=7.3, α-MeCH), 2.65 (1H, d, ³J=4.5, OH), 4.08−4.02 (1H, m,5-H), 4.08 (1H, dd, ²J=−11.6, ³J=3.5, 6-H_(A)), 4.27 (1H, dd, ²J=−11.6,³J=6.1, 6-H_(B)), 4.37 (1H, d, ³J=7.6,4-H), 4.93 (1H, q, ³J=7.3, α-CH).¹³C (100 MHz) in CDCl₃: δ13.32, 27.98, 55.25, 64.62, 70.52, 75.25,82.79, 167.33, 168.37. IR (film), cm⁻¹: 3436, 2981, 2116, 1739, 1685,1256, 1158. MS (CI, isobutane), m/Z: 287 (M⁺+1). Anal. Calcd forC₁₁H₁₈N₄O₅: C, 46.2; H, 6.3; N, 19.6. Found: C, 46.4; H, 6.3; N, 19.4%.

[0176](αR,4S,5R)-2-(α-Carboxyethyl)-4-azido-5-hydroxy-1,2-oxazinan-3-one (12).To a cooled (0˜−5° C.) stirred solution of the tert-butyl ester 11(0.469 g, 1.64 mmol) in CH₂Cl₂ (8 ml), a precooled (ca. 0° C.)trifluoroacetic acid (6.5 mL) was added portionwise. The reactionmixture was kept for 1.5 hours at room temperature and was evaporated invacuo to dryness. Traces of trifluoroacetic acid were removed byrepeated addition and evaporation of dry ether (3×15 mL). After dryingin vacuo, 0.373 g (99%) of the product 12 was obtained as a yellowishsemisolid. ¹H NMR (400 MHz) in CDCl₃ (J, Hz): δ1.58 (3H, d, ³J=7.4,α-MeCH), 4.10−4.06 (1H, m, 5-H), 4.13 (1H, dd, ²J=−11.7,³J=^(4.1 , 6)-H_(A)), 4.31 (1H, dd, ²J=−11.7, ³J=5.9, 6-H_(B)), 4.43(1H, d, ³J=7.4, 4-H), H), 5.12 (1H, q, ³J=7.4, α-CH), 6.51 (2H, br. s,OH, COOH). ¹³C (100 MHz) in CD₃CN: δ13.51, 54.74, 65.75, 71.38, 78.21,167.55, 170.91. IR (film), cm⁻¹: 3456, 2959, 2125, 1746, 1654, 1284. MS(CI, isobutane), m/Z: 231 (M⁺+1). The compound was used in the nextreaction without further purification.

[0177](αR,4S,5R)-2-(α-Carboxyethyl)-4-amino-5-hydroxy-1,2-oxazinan-3-one (13).A suspension of 5% Pd-C (0.175 g) in EtOAc (10 mL) was stirred for 1.5hours under H₂ atmosphere and a solution of the azido alcohol 12 (0.373g, 1.621 mmol) in EtOAc (15 ml) was added portionwise. The reactionmixture was stirred for 17 hours under H₂ and was evaporated to dryness.The residue was washed with dry ether (3×15 mL) and the product wasextracted from the dried residue with hot (ca. 70° C.) water (30 mL).The aqueous solution was filtered through a paper filter and the blacksolid was washed with hot water (3×10 mL). Evaporation of the filtratein vacuo provided 0.296 g (90%) of the product 13 as a white solid withmp 200-202° C. (decomp.). ¹H NMR (400 MHz) in CD₃OD+1 drop of CF₃CO₂H(J, Hz): δ1.52 (3H, d, ³J=7.4, α-MeCH), 4.08 (1H, dd, ²J=−11.7, ³J=2.5,6-H_(A)), 4.18−4.16 (2H, m, 4-H, 5-H), 4.46−4.41 (1H, dd, ²J=−11.7,³J=6.4, 6-H_(B)), 4.98 (1H, q, ³J=7.4, α-CH). ¹³C (100 MHz) in D₂O:δ17.62, 59.62, 61.42, 73.69, 82.48, 169.06, 180.86. IR (KBr), cm⁻¹:3158, 1684, 1599, 1568, 1400. MS (CI, isobutane), m/Z: 205 (M⁺+1). Anal.Calcd for C₇H₁₂N₂O₅+0.5 H₂O: C, 39.4; H, 6.1; N, 13.1. Found: C, 39.3;H, 6.2; N, 13.2%.

[0178](αR,4S,5R)-2-(α-Carboxyethyl)-4-phenylacetylamino-5-hydroxy-1,2-oxazinan-3-one(14). To a solution of the amino acid 13 (0.290 g, 1.36 mmol) and NaHCO₃(0.343 g, 4.08 mmol) in water (7 mL), a solution of phenylacetylchloride (0.191 g, 1.24 mmol) in abs. MeCN (3 mL) was added dropwisewith cooling (−5˜−6° C.) and stirring. The reaction mixture was stirredfor 2 h at 0˜−2° C. and for 1 hour at room temperature and wasconcentrated in vacuo to a volume of circa 5 mL. The solution was cooledto ca. 5° C. and adjusted to pH 2-3 with 1M aqueous HCl. (2.8 mL). Thewhite precipitate was filtered off, washed with water and dried in vacuoto afford 0.364 g (91%) of the product 14 as a white solid with mp194-195° C. (decomp.) (from iso-PrOH). ¹H NMR (400 MHz) in CD₃OD (J,Hz): δ1.47 (3H, d, ³J=7.3, α-MeCH), 3.62 (1H, d, ²J=−14.8, CH_(A)Ph),3.67 (1H, d, ²J=−14.8, CH_(B)Ph), 4.06 (1H, dd, ²J=−11.4, ³J=3.7,6-H_(A)), 4.12−4.06 (1H, m, 5-H), 4.33 (1H, dd, ²J=−11.4, ³J=5.6,6-H_(B)), 4.66 (1H, d, ³J=8.0, 4-H), 4.96 (1H, q, ³J=7.3, α-CH),7.38−7.17 (5H, m, Ph). ¹³C (100 MHz) in CD₃OD: δ1371, 43.63, 55.70,57.54, 71.84, 79.10, 127.85, 129.51, 130.37, 136.63, 169.61, 172.71,174.89. IR (KBr), cm⁻¹: 3361, 2913, 1729, 1666, 1630, 1542, 1440, 1233.MS (CI, isobutane), m/Z: 323 (M⁺+1). Anal. Calcd for C₁₅H₁₈N₂O₆: C,55.9; H, 5.6; N, 8.7. Found: C, 55.7; H, 5.7; N, 8.6%.

EXAMPLE 4

[0179] Bioassay of Oxazinones

[0180] Samples of 2-carboxymethyl-5-hydroxy-1,2-oxazin-3-one andpenicillin were applied to a filter disc in the amounts indicated inTable 1, below. The discs were applied to agar plates seeded withMicrococcus luteus, and the plates were incubated over night at 37° C.The results are summarized in the following Table 1: TABLE 1 CompoundWeight (micrograms) Zone Size (cm) Water Blank 0 Penicillin G 0.1 1.62-carboxymethyl-5-hydroxy-1,2- 100 1.1 oxazin-3-one2-carboxymethyl-5-hydroxy-1,2- 1000 2.0 oxazin-3-one

[0181] Samples of 2-[2-carboxypropyl]-5-hydroxy-1,2-oxazin-3-one anddesacetoxycephalosporin G were applied to a filter disc in the amountsindicated in Table 2, below. The discs were applied to agar platesseeded with Micrococcus luteus, and the plates were incubated overnightat 37° C. The results are summarized in the following Table 2: TABLE 2Compound Weight (micrograms) Zone Size (cm) Water Blank 0desacetoxycephalosporin G 2.5 2.5 2-[2-carboxypropyl]-5-hydroxy- 12.02.0 1,2-oxazin-3-one

[0182] With reference to Tables 1 and 2, the bioassays suggest that2-carboxymethyl-5-hydroxy-1,2-oxazin-3-one exhibits weak antibacterialactivity at least 1000 times less than that of penicillin. In contrast,2-[2-carboxypropyl]-5-hydroxy-1,2-oxazin-3-one has approximately 50times the activity of 2-carboxymethyl-5-hydroxy-1,2-oxazin-3-one.

[0183] Equivalents

[0184] Those skilled in the art will recognize, or be able to ascertainusing no more than routine experimentation, many equivalents to thespecific embodiments and methods described herein. Such equivalents areintended to be encompassed by the scope of the following claims.

[0185] All patents, patent applications, and literature references citedherein are hereby expressly incorporated by reference.

1. An olefinic oxazinone of the formula (I):

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are substituting moieties; R₇ is hydrogen, a protecting moiety,or a prodrug moiety, and acceptable salts and esters thereof.
 2. Theolefinic oxazinone of claim 1, wherein said amino acid side chainmimicking moiety is a side chain of an amino acid.
 3. The olefinicoxazinone of claim 2, wherein said amino acid is selected from the groupconsisting of alanine, valine, leucine, isoleucine, phenylalanine,tryptophan, and methionine.
 4. The olefinic oxazinone of claim 2,wherein said amino acid is selected from the group consisting ofglycine, serine, threonine, cysteine, tyrosine, asparagine, andglutamine.
 5. The olefinic oxazinone of claim 2, wherein said amino acidis selected from the group consisting of aspartic acid, glutamic acid,lysine, arginine, and histidine.
 6. The olefinic oxazinone of claim 3,wherein said amino acid is alanine.
 7. The olefinic oxazinone of claim1, wherein said amino acid mimicking moiety is alkyl.
 8. The olefinicoxazinone of claim 7, wherein said alkyl is lower alkyl.
 9. The olefinicoxazinone of claim 8, wherein said alkyl is methyl, ethyl, i-propyl,n-propyl, i-butyl, n-butyl, t-butyl, pentyl, or hexyl.
 10. The olefinicoxazinone of claim 9, wherein said alkyl is substituted with one or moresubstituents.
 11. The olefinic oxazinone of claim 1, wherein said aminoacid side chain mimicking moiety is alkenyl, alkynyl, carbonyl, aralkylor aryl.
 12. The olefinic oxazinone of claim 11, wherein said amino acidside chain mimicking moiety is aryl.
 13. The olefinic oxazinone of claim12, wherein said amino acid side chain mimicking moiety is substitutedor unsubstituted phenyl.
 14. The olefinic oxazinone of claim 1, whereinthe * carbon has an S configuration.
 15. The olefinic oxazinone of claim1, wherein the * carbon has an R configuration.
 16. The olefinicoxazinone of claim 1, wherein R₇ is hydrogen.
 17. The olefinic oxazinoneof claim 1, wherein R₄ is hydrogen or lower alkyl.
 18. The olefinicoxazinone of claim 1, wherein R₆ is hydrogen or lower alkyl.
 19. Theolefinic oxazinone of claim 1, wherein R₈ is hydrogen.
 20. The olefinicoxazinone of claim 1, wherein R₉ is hydrogen.
 21. A method forsynthesizing oxazinones of the formula (II):

wherein: R₁ is an amino acid side chain mimicking moiety; R₂ is halogen,OH, SH, NH₂, NHCOR₃, or an electronegative moiety; R₃ is anantibacterial substituent; R₄, R₈ and R₉ are each independently selectedsubstituting moieties; R₅ is OH, NH₂, NHCOR₃, or an electronegativemoiety; and R₆ is a substituting moiety or the oxygen of a carbonylgroup when taken together with R₅; R₇ is hydrogen, a protecting moiety,or a prodrug moiety, and pharmaceutically acceptable salts and estersthereof, comprising: contacting an olefinic oxazinone with aderivatizing agent, under appropriate conditions such that an oxazinoneis synthesized, wherein said olefinic oxazinone is of the formula (I):

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈ andR₉ are each independently selected substituting moieties; R₇ ishydrogen, a protecting moiety, or a prodrug moiety, and pharmaceuticallyacceptable salts and esters thereof.
 22. The method of claim 21, whereinsaid oxazinone has the following stereochemistry:


23. The method of claim 21, wherein said oxazinone has the followingstereochemistry:


24. The method of claim 21, wherein said oxazinone has the followingstereochemistry:


25. The method of claim 21, wherein said oxazinone has the followingstereochemistry:


26. The method of claim 21, further comprising the formation of anepoxide oxazinone intermediate.
 27. The method of claim 22, wherein saidepoxide oxazinone intermediate is formed stereoselectively.
 28. Themethod of claim 22 or 23, wherein said oxazinone is formed from saidepoxide oxazinone intermediate by contacting said epoxide oxazinone witha epoxide opening agent.
 29. The method of claim 21, wherein saidderivatizing agent selectively aminohydroxylates said olefinicoxazinone.
 30. The method of claim 21, wherein said derivatizing agentselectively dihydroxylates said olefinic oxazinone.
 31. The method ofclaim 21, wherein said derivatizing agent selectively halogenates saidolefinic oxazinone intermediate.
 32. The method of claim 31, whereinsaid halogenation is a fluorination.
 33. An epoxide oxazinone of theformula (III):

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are each independently selected substituting moieties; R₇ ishydrogen, a protecting moiety, or a prodrug moiety, and acceptable saltsand esters thereof
 34. An epoxide oxazinone of the formula (IV):

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are each independently selected substituting moieties; R₇ ishydrogen, a protecting moiety, or a prodrug moiety, and acceptable saltsand esters thereof.
 35. A method for the synthesis of an olefinicoxazinone comprising: contacting a diolefin with a cyclization catalystunder appropriate conditions, such that an olefinic oxazinone is formed,wherein said diolefin is of formula (V):

and wherein said olefinic oxazinone is of the formula:

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are substituting moieties; R₇ is hydrogen, a protecting moiety,or a prodrug moiety, and acceptable salts and esters thereof.
 36. Themethod of claim 35, wherein said cyclization catalyst isbis(tricyclohexylphosphine) benzylidene ruthenium dichloride.
 37. Themethod of claim 35, wherein said appropriate conditions comprise anon-polar solvent.
 38. The method of claim 37, wherein said non-polarsolvent is methylene chloride or benzene.
 39. A method for the synthesisof an olefinic oxazinone comprising: treating an olefinic triphenylphosphine salt with ozone under appropriate conditions, such that anolefinic oxazinone is formed, wherein said olefinic triphenyl phosphinesalt is of formula (VI)

and wherein said olefinic oxazinone is of the formula (I):

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are substituting moieties; R₇ is hydrogen, a protecting moiety,or a prodrug moiety, and acceptable salts and esters thereof.
 40. Themethod of claim 39, wherein said olefinic triphenyl phosphine salt is atriflic acid salt.
 41. A method for treating a bacterial associatedstate in a subject, comprising administering to said subject aneffective amount of an olefinic oxazinone, such that the subject istreated for said bacterial associated state, and wherein said olefinicoxazinone is of the formula:

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are substituting moieties; R₇ is hydrogen, a protecting moiety,or a prodrug moiety, and acceptable salts and esters thereof.
 42. Amethod for treating a bacterial associated state in a subject,comprising administering to said subject an effective amount of anepoxide oxazinone, such that the subject is treated for said bacterialassociated state and wherein said epoxide oxazinone is of the formula(III):

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are each independently selected substituting moieties; R₇ ishydrogen, a protecting moiety, or a prodrug moiety, and acceptable saltsand esters thereof.
 43. A method for treating a bacterial associatedstate in a subject, comprising administering to said subject aneffective amount of an epoxide oxazinone, such that the subject istreated for said bacterial associated state, and wherein said epoxideoxazinone is of the formula:

wherein: R₁ is an amino acid side chain mimicking moiety; R₄, R₆, R₈,and R₉ are each independently selected substituting moieties; R₇ ishydrogen, a protecting moiety, or a prodrug moiety, and acceptable saltsand esters thereof.
 44. A pharmaceutical composition, comprising aneffective amount compound of claims 1, 33, or 34 and a pharmaceuticallyacceptable carrier.
 45. The pharmaceutical composition of claim 44,wherein said effective amount is effective to treat a bacterialassociated state in a subject.